Information on EC 4.3.3.7 - 4-hydroxy-tetrahydrodipicolinate synthase

New: Word Map on EC 4.3.3.7
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea

EC NUMBER
COMMENTARY
4.3.3.7
-
RECOMMENDED NAME
GeneOntology No.
4-hydroxy-tetrahydrodipicolinate synthase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
pyruvate + L-aspartate-4-semialdehyde = (2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O
show the reaction diagram
-
-
-
-
pyruvate + L-aspartate-4-semialdehyde = (2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O
show the reaction diagram
pyruvate binds to the enzyme first by forming a Schiff base with the epsilon-amino group of Lys161. After release of the first water molecule L-aspartate-4-semialdehyde binds to the active site and the condensation reaction to form 2,3-dihydrodipicolinate takes place
-
pyruvate + L-aspartate-4-semialdehyde = (2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O
show the reaction diagram
the reaction is initiated by an nucleophilic attack of the epsilon-amino group of active site lysine to the C2 atom of pyruvate with subsequent imine formation. Tautomerization of the imine to the enamine creates the nucleophile necessary for the attack of the aldehyde group of L-aspartate semialdehyde. Cyclization of the substrate coupled with transamination leads to stepwise detachment from the active site
-
pyruvate + L-aspartate-4-semialdehyde = (2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O
show the reaction diagram
ping-pong mechanism, pyruvate binds as a Schiff base to Lys161. After aldol reaction with L-aspartate-4-semialdehyde and transamination the product is released
-
pyruvate + L-aspartate-4-semialdehyde = (2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O
show the reaction diagram
ping-pong kinetic and catalytic mechanism involves a catalytic triad which consists of Tyr133, Thr44, and Tyr107, overview
-
pyruvate + L-aspartate-4-semialdehyde = (2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O
show the reaction diagram
reaction mechanism including Schiff base formation, tautomerization, and cyclization, overview
-
pyruvate + L-aspartate-4-semialdehyde = (2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O
show the reaction diagram
ping-pong kinetic reaction mechanism
-
pyruvate + L-aspartate-4-semialdehyde = (2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O
show the reaction diagram
ping-pong mechanism, in which pyruvate binds as a Schiff base to an active-site lysine residue (Lys162 in Agrobacterium tumefaciens)
Q8UGL3
pyruvate + L-aspartate-4-semialdehyde = (2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O
show the reaction diagram
reaction steps, overview
O67216
pyruvate + L-aspartate-4-semialdehyde = (2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O
show the reaction diagram
the enzyme-catalyzed reaction is initiated by condensation of pyruvate with an active site lysine residue (Lys161 in Escherichia coli DHDPS) forming a Schiff base, ping-pong kinetic reaction mechanism via enamine intermediate, overview
-
pyruvate + L-aspartate-4-semialdehyde = (2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O
show the reaction diagram
the reaction proceeds via a ping-pong kinetic mechanism in which pyruvate binds and forms a Schiff base to an active-site lysine residue (Lys184 in Vitis vinifera enzyme). L-aspartate-4-semialdehyde then reacts with the resultant enamine and following cyclization forms the product (2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate
D7U7T8
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
elimination
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biosynthesis of antibiotics
-
-
Biosynthesis of secondary metabolites
-
-
L-lysine biosynthesis I
-
-
L-lysine biosynthesis II
-
-
L-lysine biosynthesis III
-
-
L-lysine biosynthesis VI
-
-
Lysine biosynthesis
-
-
lysine metabolism
-
-
Metabolic pathways
-
-
Microbial metabolism in diverse environments
-
-
Monobactam biosynthesis
-
-
SYSTEMATIC NAME
IUBMB Comments
L-aspartate-4-semialdehyde hydro-lyase [adding pyruvate and cyclizing; (4S)-4-hydroxy-2,3,4,5-tetrahydro-(2S)-dipicolinate-forming]
Studies of the enzyme from the bacterium Escherichia coli have shown that the reaction can be divided into three consecutive steps: Schiff base formation between pyruvate and an active-site lysine, the addition of L-aspartate-semialdehyde, and finally transimination leading to cyclization with simultaneous dissociation of the product.
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
DHDPS
-
-
-
-
dihydrodipicolinic acid synthase
-
-
-
-
dihydropicolinate synthetase
-
-
-
-
pyruvate-aspartic semialdehyde condensing enzyme
-
-
-
-
synthase, dihydrodipicolinate
-
-
-
-
VEG81
-
-
-
-
Vegetative protein 81
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
9055-59-8
-
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
dapA10; gene dapA10
UniProt
Manually annotated by BRENDA team
dapA1; gene dapA1
UniProt
Manually annotated by BRENDA team
dapA2; genes dapA2
UniProt
Manually annotated by BRENDA team
dapA3; gene dapA3
UniProt
Manually annotated by BRENDA team
dapA4; gene dapA4
UniProt
Manually annotated by BRENDA team
dapA5; gene dapA5
UniProt
Manually annotated by BRENDA team
dapA6; gene dapA6
UniProt
Manually annotated by BRENDA team
dapA7; gene dapA7
UniProt
Manually annotated by BRENDA team
dapA8; gene dapA8
UniProt
Manually annotated by BRENDA team
dapA9; gene dapA9
UniProt
Manually annotated by BRENDA team
gene dapA or Aq_1143
UniProt
Manually annotated by BRENDA team
Sterne strain
UniProt
Manually annotated by BRENDA team
Bacillus anthracis Sterne
Sterne strain
UniProt
Manually annotated by BRENDA team
cabbage
-
-
Manually annotated by BRENDA team
Cucurbita sp.
-
-
-
Manually annotated by BRENDA team
strain XL1-Blue expressing the enzyme from a plasmid, gene dapA
-
-
Manually annotated by BRENDA team
Escherichia coli MG1655
gene dapA
-
-
Manually annotated by BRENDA team
Escherichia coli XL1-Blue
strain XL1-Blue expressing the enzyme from a plasmid, gene dapA
-
-
Manually annotated by BRENDA team
barley
-
-
Manually annotated by BRENDA team
four isozymes, MtDHDPS, MtDHDPS2, MtDHDPS3, and MtDHDPS4
-
-
Manually annotated by BRENDA team
Medicago truncatula Jemalong 2HA
four isozymes, MtDHDPS, MtDHDPS2, MtDHDPS3, and MtDHDPS4
-
-
Manually annotated by BRENDA team
strain MC58
UniProt
Manually annotated by BRENDA team
tobacco
-
-
Manually annotated by BRENDA team
no activity in Homo sapiens
-
-
-
Manually annotated by BRENDA team
Phaseolus sp.
bean
-
-
Manually annotated by BRENDA team
gene dapA or PA1010
UniProt
Manually annotated by BRENDA team
gene dapA
-
-
Manually annotated by BRENDA team
Shewanella benthica ATCC3392
gene dapA
-
-
Manually annotated by BRENDA team
strain L5-30
-
-
Manually annotated by BRENDA team
Sinorhizobium meliloti L5-30
L5-30
SwissProt
Manually annotated by BRENDA team
Sinorhizobium meliloti L5-30
strain L5-30
-
-
Manually annotated by BRENDA team
potato, increased levels in transgenic plants containing the Escherichia coli gene
-
-
Manually annotated by BRENDA team
a methicillin-resistant strain
-
-
Manually annotated by BRENDA team
methicillin-resistant
-
-
Manually annotated by BRENDA team
methicillin-resistent MRSA252 strain
SwissProt
Manually annotated by BRENDA team
strain COL
SwissProt
Manually annotated by BRENDA team
Staphylococcus aureus COL
strain COL
SwissProt
Manually annotated by BRENDA team
Staphylococcus aureus MRSA252
methicillin-resistent MRSA252 strain
SwissProt
Manually annotated by BRENDA team
strain OXC141
-
-
Manually annotated by BRENDA team
gene dapA
Uniprot
Manually annotated by BRENDA team
corn
-
-
Manually annotated by BRENDA team
wild-type and four high-lysine maize mutants (Oh43o1, Oh43o2, Oh43fl1 and Oh43fl2)
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
Q9I4W3
enzyme mutants with deleted dapA gene are viable and able to grow in a mouse lung infection model
metabolism
-
sequential production of dihydrodipicolinate and dipicolinic acid appears to be catalysed by DHDPA synthase followed by an electron transfer flavoprotein, EtfA from Clostridium perfringens. Spontaneous dipicolinic acid formation in the presence of high concentrations of DapA
metabolism
-
DHDPS is an oligomeric enzyme that catalyzes the first committed step of the lysine biosynthesis pathway in plants and bacteria, which yields essential building blocks for cell-wall and protein synthesis
metabolism
-
first enzyme unique to the diaminopimelate pathway of lysine biosynthesis
metabolism
-
dihydrodipicolinate synthase is a key enzyme in the lysine biosynthesis pathway that catalyzes the condensation of pyruvate and aspartate semi-aldehyde
metabolism
-
feedback regulation of the enzyme is directly correlated to L-lysine production
metabolism
-
lysine biosynthesis in plants is tightly regulated by feedback inhibition of the end product on dihydrodipicolinate synthase, the first enzyme of the lysine-specific branch
metabolism
D7U7T8
lysine biosynthesis in plants is tightly regulated by feedback inhibition of the end product on dihydrodipicolinate synthase, the first enzyme of the lysine-specific branch
metabolism
-
the enzyme catalyses the first committed step in the lysine biosynthesis pathway, the condensation of pyruvate and (S)-aspartate semialdehyde to form (2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid
metabolism
-
the enzyme catalyses the first committed step in the lysine biosynthesis pathway, the condensation of pyruvate and (S)-aspartate semialdehyde to form (4S)-4-hydroxy-2,3,4,5-tetrahydro-(2S)-dipicolinic acid
metabolism
-
the enzyme catalyses the first committed step in the lysine biosynthesis pathway, the condensation of pyruvate and (S)-aspartate semialdehyde to form (4S)-4-hydroxy-2,3,4,5-tetrahydro-(2S)-dipicolinic acid
metabolism
Q8UGL3
the enzyme catalyses the first committed step in the lysine biosynthesis pathway, the condensation of pyruvate and (S)-aspartate semialdehyde to form (4S)-4-hydroxy-2,3,4,5-tetrahydro-(2S)-dipicolinic acid
metabolism
O67216
the enzyme catalyses the first committed step in the lysine biosynthesis pathway, the condensation of pyruvate and (S)-aspartate semialdehyde to form (4S)-4-hydroxy-2,3,4,5-tetrahydro-(2S)-dipicolinic acid
metabolism
Q9FVC8
the enzyme catalyzes the branch-point reaction in the biosynthetic pathway leading to meso-diaminopimelate and (S)-lysine in plants and bacteria: condensation of (S)-aspartate-4-semialdehyde and pyruvate to form (4S)-4-hydroxy-2,3,4,5-tetrahydro-(2S)-dipicolinic acid. There are four variants of the meso-diaminopimelate/(S)-lysine pathway, they all share the same enzymatic steps for the synthesis of tetrahydrodipicolinate from aspartate, which includes the reaction catalyzed by the enzyme, overview
metabolism
Q9I4W3
the enzyme catalyzes the first step in the diaminopimelic acid pathway of lysine biosynthesis
metabolism
Shewanella benthica ATCC3392
-
DHDPS is an oligomeric enzyme that catalyzes the first committed step of the lysine biosynthesis pathway in plants and bacteria, which yields essential building blocks for cell-wall and protein synthesis
-
metabolism
Escherichia coli MG1655
-
feedback regulation of the enzyme is directly correlated to L-lysine production
-
physiological function
-
complementation of the auxotrophy of Escherichia coli XL1-Blue KanRDELTAdapA cells only with the plasmid pUCX:dapA encoding wild-type DHDPS
physiological function
Q1PDD5
DHDPS can recover the DHDPS-deleted mutant of Escherichia coli
physiological function
-
DHDPS catalyses a branch point reaction the condensation of pyruvate and (S)-aspartate beta-semialdehyde to form an unstable product, (4S)-4-hydroxy-2,3,4,5-tetrahydro-(2S)-dipicolinic acid, which is ultimately advanced to the final metabolites (S)-lysine and meso-diaminopimelate
physiological function
O67216
the enzyme catalyzes a rate-limiting step in the (S)-lysine biosynthetic pathway, which is regulated by a feedback mechanism through lysine
metabolism
Medicago truncatula Jemalong 2HA
-
lysine biosynthesis in plants is tightly regulated by feedback inhibition of the end product on dihydrodipicolinate synthase, the first enzyme of the lysine-specific branch
-
additional information
-
monomeric species exhibit an enhanced propensity for aggregation and inactivation, indicating that whilst the oligomerization is not an intrinsic criterion for catalysis, higher oligomeric forms may benefit from both increased catalytic efficiency and diminished aggregation propensity
additional information
-
pyruvate binding occurs near the large interface of DHDPS, and is likely, therefore, to stabilize this solvent-accessible face, which favors the formation of a dimer rather than a monomer. For the DELTAAsp168/Arg237 and DELTAAsp168/Asp171 DHDPS variants addition of pyruvate shifts the equilibrium from primarily monomer to favor almost exclusively dimers. On the other hand, for the DELTAAsp168 DHDPS variant, the monomer-tetramer equilibrium shifts from primarily monomer to primarily tetramer on addition of pyruvate
additional information
-
the tetrameric structure is not essential for activity in DHDPS from Mycobacterium tuberculosis
additional information
-
optimal activity is achieved by minimizing the inherent dimer flexibility using buttressing two dimers together in the case of the Escherichia coli tetrameric enzyme, active site structure simulations, overview
additional information
-
optimal activity is achieved by minimizing the inherent dimer flexibility using strengthening and extending the dimer interface in the dimeric Staphylococcus aurus MRSA strain enzyme, active site structure simulations, overview
additional information
-
structure comparison with the enzyme from Corynebacterium glutamicum
additional information
-
structure comparison with the enzyme from Escherichia coli
additional information
Q9I4W3
structure-based sequence alignments, based on the DapA crystal structure, reveal the presence of two homologues, PA0223 and PA4188, in Pseudomonas aeruginosa that can substitute for DapA in the PAO1DELTAdapA mutant. In vitro experiments using recombinant PA0223 protein do not detect any DapA activity
additional information
-
the catalytic site of DHDPS is situated at the C-terminal end of the TIM barrel where the pyruvate-binding residue, Lys161, lies in a solvent-accessible cleft with Arg138 capping the binding site. One-half of the active site is blocked by binding interactions of another monomer
additional information
-
the catalytic triad, consisting of Tyr133, Thr44, and Tyr107, acts as a proton relay to transfer protons to and from the active site via a water-filled channel leading to bulk solvent
additional information
O67216
the enzyme has unique disulfide linkage which is critical for the stability of the enzyme tetramer, but is not conserved in homologous enzymes, molecular dynamics simulation of the native structure of the enzyme tetramer and dimeric units containing complexes of enzyme-pyruvate or enzyme-lysine, enzyme structure comparisons and ligand docking analysis, overview
additional information
-
structure comparison with the enzyme from Escherichia coli
-
additional information
Escherichia coli MG1655
-
structure comparison with the enzyme from Corynebacterium glutamicum
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
P0A6L2
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
Q5HG25
-
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
Q8RBI5
-
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
P0A6L2
branch point reaction in the biosynthesis of lysine
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
first step in the biosynthesis of lysine, overview
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
Q9X1K9
catalyses the branch point in lysine biosynthesis
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
biosynthesis of (S)-lysine and meso-diaminopimelate
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
Q5HG25
biosynthesis of (S)-lysine and meso-diaminopimelate, compounds of bacterial cell walls
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
Q8RBI5
characterization of thermostable enzyme features, substrate specificity tested, activity measurements at elevated temperature of 60C
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
diketopimelic acid derivatives designed as mimics of the acyclic enzyme-bound condensation product of (S)-aspartate 4-semialdehyde and pyruvate, inhibition analysis
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
mechanistic insight into catalysis, structural features, evolution of quaternary structure
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
reaction mechanism and active site analyzed, inhibition by the substrate analog beta-hydroxypyruvate
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
role of Tyr107 residue, located at the tight-dimer interface between two monomers, participates in a catalytic triad of residues during catalysis
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
Staphylococcus aureus MRSA252
-
mechanistic insight into catalysis, structural features, evolution of quaternary structure
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
Escherichia coli XL1-Blue
-
first step in the biosynthesis of lysine, overview
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
Staphylococcus aureus COL
Q5HG25
biosynthesis of (S)-lysine and meso-diaminopimelate, compounds of bacterial cell walls
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
-
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
-
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
-
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
-
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
-
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
-
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
Q9I4W3
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
A9CFV4, A9CGZ4, A9CHR2, A9CL94, A9CL97, Q7CU96, Q7D0E8, Q7D313, Q7D3Z9, Q8UGL3
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
Q8UGL3
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
Q9FVC8
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
O67216
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
-
the enzyme-catalyzed reaction is initiated by condensation of pyruvate with an active site Lys161 forming a Schiff base. Subsequent tautomerization to the enamine and aldol-type reaction with (S)-aspartate-4-semialdehyde then generates an acyclic enzyme-bound intermediate. Transimination of the acyclic intermediate yields the cyclic alcohol (4S)-4-hydroxy-2,3,4,5-tetrahydro-(2S)-dipicolinic acid, with simultaneous release of the active site lysine residue
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
-
(S)-aspartate-4-semialdehyde binds cooperatively in the presence of (S)-lysine with an average cooperativity coefficient n = 1.3, and the cooperativity of binding increases at near-KM concentrations of pyruvate with an average cooperativity coefficient n = 1.0
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
A9CFV4, A9CGZ4, A9CHR2, A9CL94, A9CL97, Q7CU96, Q7D0E8, Q7D313, Q7D3Z9, Q8UGL3
bi-bi ping-pong substrate model
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
Corynebacterium glutamicum ATCC 13032, Escherichia coli MG1655
-
-
-
-
?
L-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
-
?
L-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
-
?
L-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
Q1PDD5
-
-
-
?
L-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
branch poit of (S)-lysine biosynthesis
-
-
?
L-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
isothermal titration calorimetry (ITC) to study Schiff base formation between an enzyme and its substrate, pyruvate condenses with an active site lysine residue (Lys161 in MosA)
-
-
?
L-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
P19808
lysine insensitivity of dihydrodipicolinate synthase analyzed, catalytic lysine residue forms a Schiff base adduct with pyruvate, active site lysine residues (K176a, K176b)
-
-
?
L-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
substrate protection and inhibition experiments
-
-
?
L-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
Sinorhizobium meliloti L5-30
-
isothermal titration calorimetry (ITC) to study Schiff base formation between an enzyme and its substrate, pyruvate condenses with an active site lysine residue (Lys161 in MosA)
-
-
?
L-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + 2 H2O
show the reaction diagram
-
-
-
-
?
L-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + 2 H2O
show the reaction diagram
-
-
-
-
?
L-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + 2 H2O
show the reaction diagram
Q81WN7
-
-
-
?
L-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + 2 H2O
show the reaction diagram
-
-
-
-
?
L-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + 2 H2O
show the reaction diagram
-
-
-
-
?
L-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + 2 H2O
show the reaction diagram
-
active sites are similiar to Bacillus anthracis DHDPS
-
-
?
L-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + 2 H2O
show the reaction diagram
-
active sites are similiar to Escherichia coli DHDPS
-
-
?
L-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + 2 H2O
show the reaction diagram
-
condensation reaction between both substrates via the formation of a Schiff base intermediate between pyruvate and the absolutely conserved active-site lysine. Although lysine 161 is important in the wild-type DHDPS-catalysed reaction, it is not absolutely essential for catalysis
-
-
?
L-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + 2 H2O
show the reaction diagram
-
pyruvate is a weak binder (0.023 mM) and L-aspartate 4-semialdehyde does not interact with the enzyme in the absence of a Schiff-base with pyruvate. Lys161 plays a crucial role in providing the thermodynamic force for the association of pyruvate with the DHDPS active site
-
-
?
L-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + 2 H2O
show the reaction diagram
Q9JZR4
slightly higher affinity for L-aspartate 4-semialdehyde and somewhat lower affinity for pyruvate than the Escherichia coli enzyme, whereby the catalytic activity is similar
-
-
?
L-aspartate 4-semialdehyde + pyruvate
(S)-2,3-dihydropyridine-2,6-dicarboxylate + 2 H2O
show the reaction diagram
-
-
-
-
?
L-aspartate 4-semialdehyde + pyruvate
(S)-2,3-dihydropyridine-2,6-dicarboxylate + 2 H2O
show the reaction diagram
-
-
-
-
?
L-aspartate 4-semialdehyde + pyruvate
(S)-2,3-dihydropyridine-2,6-dicarboxylate + 2 H2O
show the reaction diagram
-
-
-
-
?
L-aspartate 4-semialdehyde + pyruvate
(S)-2,3-dihydropyridine-2,6-dicarboxylate + 2 H2O
show the reaction diagram
Q9I4W3
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
Cucurbita sp., Phaseolus sp., Brassica sp.
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
(4S)-4-hydroxy-2,3,4,5-terahydro-(2S)-dipicolinic acid is the initial but instable product
?
L-aspartate-4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
D-isomer inactive
-
?
L-aspartate-4-semialdehyde + pyruvate
?
show the reaction diagram
-
-
-
-
-
L-aspartate-4-semialdehyde + pyruvate
?
show the reaction diagram
-
-
-
-
-
L-aspartate-4-semialdehyde + pyruvate
?
show the reaction diagram
-
biosynthesis of lysine via the diaminopimelate pathway
-
-
-
L-aspartate-4-semialdehyde + pyruvate
?
show the reaction diagram
-
synthesis of the precursor of dipicolinic acid which plays a key role in bacterial sporulation process
-
-
-
pyruvate + L-aspartate-4-semialdehyde
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O
show the reaction diagram
-
-
-
-
?
pyruvate + L-aspartate-4-semialdehyde
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O
show the reaction diagram
-
-
-
-
?
pyruvate + L-aspartate-4-semialdehyde
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O
show the reaction diagram
D7U7T8
-
-
-
?
pyruvate + L-aspartate-4-semialdehyde
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O
show the reaction diagram
Medicago truncatula Jemalong 2HA
-
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
?
show the reaction diagram
-
first enzyme in the meso-diaminopimelate and L-lysine branch of the aspartate pathway
-
-
-
additional information
?
-
-
no reaction with oxaloacetic acid, phosphoenolpyruvate, glutamic semialdehyde, N-acetylaspartic semialdehyde, succinic semialdehyde
-
-
-
additional information
?
-
-
no activity with (R)-aspartate 4-semialdehyde
-
?
additional information
?
-
Q9I4W3
the active site is formed by residues Thr44, Tyr107 and Tyr133, stereochemically suitable for catalytic function
-
-
-
additional information
?
-
-
the active site of the monomer is well conserved, with most active-site residues in the same conformation
-
-
-
additional information
?
-
-
the quaternary structure plays a significant role in substrate specificity, overview
-
-
-
additional information
?
-
-
coupled assay with dihydrodipicolinate reductase
-
-
-
additional information
?
-
D7U7T8
coupled assay with dihydrodipicolinate reductase
-
-
-
additional information
?
-
-
isozyme MtDHDPS1, followed by isozyme MtDHDPS4, exhibits the highest activity, while activity of isozyme MtDHDPS2 and especially of isozyme MtDHDPS3 is low
-
-
-
additional information
?
-
Escherichia coli XL1-Blue
-
no activity with (R)-aspartate 4-semialdehyde
-
?
additional information
?
-
Escherichia coli MG1655
-
coupled assay with dihydrodipicolinate reductase
-
-
-
additional information
?
-
Medicago truncatula Jemalong 2HA
-
isozyme MtDHDPS1, followed by isozyme MtDHDPS4, exhibits the highest activity, while activity of isozyme MtDHDPS2 and especially of isozyme MtDHDPS3 is low
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
Q5HG25
-
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
Q8RBI5
-
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
P0A6L2
branch point reaction in the biosynthesis of lysine
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
first step in the biosynthesis of lysine, overview
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
Q9X1K9
catalyses the branch point in lysine biosynthesis
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
Staphylococcus aureus MRSA252
-
-
-
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
Escherichia coli XL1-Blue
-
first step in the biosynthesis of lysine, overview
-
?
(S)-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
Staphylococcus aureus COL
Q5HG25
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
-
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
-
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
-
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
-
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
-
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
Q9I4W3
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
A9CFV4, A9CGZ4, A9CHR2, A9CL94, A9CL97, Q7CU96, Q7D0E8, Q7D313, Q7D3Z9, Q8UGL3
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
Q8UGL3
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
Q9FVC8
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
O67216
-
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
-
the enzyme-catalyzed reaction is initiated by condensation of pyruvate with an active site Lys161 forming a Schiff base. Subsequent tautomerization to the enamine and aldol-type reaction with (S)-aspartate-4-semialdehyde then generates an acyclic enzyme-bound intermediate. Transimination of the acyclic intermediate yields the cyclic alcohol (4S)-4-hydroxy-2,3,4,5-tetrahydro-(2S)-dipicolinic acid, with simultaneous release of the active site lysine residue
-
-
?
(S)-aspartate-4-semialdehyde + pyruvate
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinic acid + H2O
show the reaction diagram
Corynebacterium glutamicum ATCC 13032, Escherichia coli MG1655
-
-
-
-
?
L-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
-
?
L-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
-
-
-
-
?
L-aspartate 4-semialdehyde + pyruvate
(S)-2,3-dihydropyridine-2,6-dicarboxylate + 2 H2O
show the reaction diagram
-
-
-
-
?
L-aspartate 4-semialdehyde + pyruvate
(S)-2,3-dihydropyridine-2,6-dicarboxylate + 2 H2O
show the reaction diagram
-
-
-
-
?
L-aspartate 4-semialdehyde + pyruvate
(S)-2,3-dihydropyridine-2,6-dicarboxylate + 2 H2O
show the reaction diagram
-
-
-
-
?
L-aspartate 4-semialdehyde + pyruvate
(S)-2,3-dihydropyridine-2,6-dicarboxylate + 2 H2O
show the reaction diagram
Q9I4W3
-
-
-
?
L-aspartate 4-semialdehyde + pyruvate
dihydrodipicolinate + H2O
show the reaction diagram
Sinorhizobium meliloti L5-30
-
-
-
-
?
L-aspartate-4-semialdehyde + pyruvate
?
show the reaction diagram
-
-
-
-
-
L-aspartate-4-semialdehyde + pyruvate
?
show the reaction diagram
-
-
-
-
-
L-aspartate-4-semialdehyde + pyruvate
?
show the reaction diagram
-
biosynthesis of lysine via the diaminopimelate pathway
-
-
-
L-aspartate-4-semialdehyde + pyruvate
?
show the reaction diagram
-
synthesis of the precursor of dipicolinic acid which plays a key role in bacterial sporulation process
-
-
-
L-aspartate-4-semialdehyde + pyruvate
?
show the reaction diagram
-
first enzyme in the meso-diaminopimelate and L-lysine branch of the aspartate pathway
-
-
-
pyruvate + L-aspartate-4-semialdehyde
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O
show the reaction diagram
-
-
-
-
?
pyruvate + L-aspartate-4-semialdehyde
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O
show the reaction diagram
D7U7T8
-
-
-
?
pyruvate + L-aspartate-4-semialdehyde
(2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O
show the reaction diagram
Medicago truncatula Jemalong 2HA
-
-
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
FAD
-
produces a 5fold increase in dipicolinic acid production
FMN
-
results in a 3fold increase in dipicolinic acid production
additional information
-
neither NAD+ nor NADP+ are involved in the production of dipicolinic acid
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
no effect: Cd2+, Mg2+, Mn2+, Zn2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
(1SR,3R,5S)-1-hydroxy-3,5-bis(methoxycarbonyl)thiomorpholin-1-ium
-
30 mM, 18% inhibition
(1SS,3R,5S)-1-hydroxy-3,5-bis(methoxycarbonyl)thiomorpholin-1-ium
-
9 mM, 48% inhibition
(2E)-4-oxohept-2-enedioic acid
-
-
(2E,5E)-4-oxohepta-2,5-dienedioic acid
-
-
(2R,6S)-piperidine-2,6-dicarboxylic acid
-
IC50: 20 mM, 83% inhibition at 50 mM
(2R,6S)-piperidine-2,6-dicarboxylic acid
-
20 mM, 49% inhibition
(2R,6S)-piperidine-2,6-dicarboxylic acid
-
20 mM, 43% inhibition; 20 mM, 77% inhibition
(2R,6S)-piperidine-2,6-dicarboxylic acid
-
20 mM, 51% inhibition
(3R,5R)-thiomorpholine-3,5-dicarboxylic acid
-
50 mM, 32% inhibition
(3R,5R)-thiomorpholine-3,5-dicarboxylic acid 1,1-dioxide
-
50 mM, 11% inhibition
(3R,5S)-thiomorpholine-3,5-dicarboxylic acid 1,1-dioxide
-
50 mM, 14% inhibition
(S)-aspartate 4-semialdehyde
-
-
(S)-aspartate-4-semialdehyde
A9CFV4, A9CGZ4, A9CHR2, A9CL94, A9CL97, Q7CU96, Q7D0E8, Q7D313, Q7D3Z9, Q8UGL3
substrate inhibition
(S)-Lys
-
partial mixed inhibition with respect to pyruvate and partial non-competitive inhibition with resoect to L-aspartate 4-semialdehyde
(S)-lysine
-
mutant enzymes R138H and R138A show the same IC50 values as the wild-type enzyme, but different partial inhibition patterns
(S)-lysine
-
partial mixed inhibition with respect to its first substrate, pyruvate
(S)-lysine
-
partial mixed inhibition with respect to (S)-aspartate 4-semialdehyde, partial non-ncompetitive inhibition with respect to pyruvate in wild-type and in Y107W mutant, Y107W mutant still retains over 25% of uninhibited activity, even at high inhibitor concentrations compared to the wild-type enzyme retaining less than 10% of normal activity
(S)-lysine
-
no significant conformational change between the pyruvate-bound and (S)-lysine-bound enzyme, presence of substrate has substantial effect on the nature of enzyme-inhibitor association, solvent plays a key role in binding of inhibitor
(S)-lysine
Q8RBI5
increasing amounts of (S)-lysine (0-200 mM) added, not inhibited under physiological conditions, binding site of the allosteric inhibitor lysine appears not to be conserved
(S)-lysine
-
allosteric inhibitor
(S)-lysine
-
feedback inhibition, feedback inhibition of the Escherichia coli enzyme by lysine is successfully alleviated after substitution of the residues around the inhibitor's binding sites with those of the Corynabacterium glutamicum enzyme
(S)-lysine
-
feedback inhibition, the tetrameric enzyme has two allosteric sites, each of which binds two molecules of lysine. Lysine binds highly cooperatively, and primarily to the F form of the enzyme during the ping-pong mechanism. (S)-Lysine is an uncompetitive partial inhibitor with respect to its first substrate, pyruvate, and a mixed partial inhibitor with respect to its second substrate, (S)-aspartate-4-semialdehyde
(S)-lysine
O67216
feedback inhibitor, docking study with enzyme crystals
(S)-lysine
A9CFV4, A9CGZ4, A9CHR2, A9CL94, A9CL97, Q7CU96, Q7D0E8, Q7D313, Q7D3Z9, Q8UGL3
allosteric feedback inhibition; allosteric feedback inhibition; allosteric feedback inhibition; allosteric feedback inhibition; allosteric feedback inhibition; allosteric feedback inhibition; allosteric feedback inhibition, allosteric site modeling; allosteric feedback inhibition, allosteric site modeling; allosteric feedback inhibition, allosteric site modeling; allosteric feedback inhibition, allosteric site modeling
2,2'-(2-hydroxy-1,3-phenylene)bis(2-oxoacetic acid)
-
slow-tight inhibition
-
2,2'-benzene-1,3-diylbis(oxoacetic acid)
-
-
2,2'-benzene-1,3-diylbis(oxoacetic acid)
-
slow inhibition
2-fluoropyruvate
-
-
-
2-oxobutyrate
-
forms a Schiff base with MosA protein, competitive inhibition
2-oxobutyrate
-
competitive inhibitor of DHDPS
2-oxoglutarate
-
competitive inhibitor of DHDPS
2-oxopimelic acid
-
-
3-Bromopyruvate
-
-
3-Bromopyruvate
-
-
3-Bromopyruvate
P19808
competitive inhibition, pyruvate as varied substrate
3-bromopyruvate ethyl ester
-
1.2 mM, 85% inhibition
3-Fluoropyruvate
-
competitive inhibitor of DHDPS, and a competitive substrates
3-hydroxy-2-oxopropanoate
-
time-dependent inhibition, qualitatively followed by mass spectrometry, initial noncovalent adduct formation, followed by the slow formation of the covalent adduct
3-hydroxypyruvate
-
competitive inhibitor of DHDPS and a competitive substrate
4-oxo-1,4-dihydropyridine-2,6-dicarboxylic acid
-
i.e. chelidamic acid, IC50: 22 mM, 99% inhibition at 50 mM, uncompetitive inhibitor with respect to both substrates
4-oxo-1,4-dihydropyridine-2,6-dicarboxylic acid
-
-
4-oxo-1,4-dihydropyridine-2,6-dicarboxylic acid
-
20 mM, 73% inhibition
4-oxo-1,4-dihydropyridine-2,6-dicarboxylic acid
-
20 mM, 83% inhibition
Bromopyruvate
-
is an irreversible inhibitor of DHDPS
cis-(1SS,3R5S)-3,5-thiomorpholinedicarboxylic acid, dimethyl ester, 1-oxide
-
20 mM, 8% inhibition
cis-(1SS,3R5S)-3,5-thiomorpholinedicarboxylic acid, dimethyl ester, 1-oxide
-
20 mM, 10% inhibition
cis-piperidine-2,6-dicarboxylic acid
-
and derivatives
D-Lysine
Q8UGL3
allosteric inhibitor
diethyl (2E)-4-oxohept-2-enedioate
-
-
diethyl (2E,5E)-4-oxohepta-2,5-dienedioate
-
-
dimethyl (2R,6S)-piperidine-2,6-dicarboxylate
-
20 mM, 92% inhibition
dimethyl (2R,6S)-piperidine-2,6-dicarboxylate
-
20 mM, 24% inhibition; 20 mM, 99% inhibitione
dimethyl (2R,6S)-piperidine-2,6-dicarboxylate
-
20 mM, 58% inhibition
dimethyl (3R,5R)-thiomorpholine-3,5-dicarboxylate
-
20 mM, 12% inhibition
dimethyl (3R,5R)-thiomorpholine-3,5-dicarboxylate
-
20 mM, 19% inhibition
dimethyl (3R,5R)-thiomorpholine-3,5-dicarboxylate 1,1-dioxide
-
50 mM, 12% inhibition
dimethyl (3R,5R)-thiomorpholine-3,5-dicarboxylate 1,1-dioxide
-
20 mM, 14% inhibition
dimethyl (3R,5R)-thiomorpholine-3,5-dicarboxylate 1,1-dioxide
-
20 mM, 6% inhibition; 20 mM, 7% inhibition
dimethyl (3R,5R)-thiomorpholine-3,5-dicarboxylate 1-oxide
-
20 mM, 7% inhibition
dimethyl (3R,5S)-thiomorpholine-3,5-dicarboxylate
-
50 mM, 36% inhibition
dimethyl (3R,5S)-thiomorpholine-3,5-dicarboxylate
-
20 mM, 35% inhibition
dimethyl (3R,5S)-thiomorpholine-3,5-dicarboxylate
-
20 mM, 23% inhibition
dimethyl (3R,5S)-thiomorpholine-3,5-dicarboxylate 1,1-dioxide
-
20 mM, 19% inhibition; 20 mM, 6% inhibition
dimethyl 2,2'-benzene-1,3-diylbis[(hydroxyimino)ethanoate]
-
-
dimethyl 4-oxo-1,4-dihydropyridine-2,6-dicarboxylate
-
-
dimethyl 4-oxo-1,4-dihydropyridine-2,6-dicarboxylate
-
20 mM, 84% inhibition
dimethyl 4-oxo-1,4-dihydropyridine-2,6-dicarboxylate
-
20 mM, 88% inhibition
dimethyl chelidamate
-
IC50: 14 mM. 99% inhibition at 50 mM, noncompetitive with respect to both substrates
dimethyl piperidine-2,6-dicarboxylate
-
-
dimethyl pyridine-2,6-dicarboxylate
-
20 mM, 5% inhibition
dimethyl pyridine-2,6-dicarboxylate
-
20 mM, 13% inhibition
dimethyl-(2E,2'E)-2,2'-benzene-1,3-diylbis[(hydroxyimino)ethanoate]
-
slow inhibition
dimethyl-2,2'-(2-hydroxy-1,3-phenylene)bis(2-oxoacetate)
-
slow-tight inhibition
-
dipicolinic acid
-
1.2 mM, 50% inhibition
dipicolinic acid
-
1 mM, 50% inhibition
dipicolinic acid
-
IC50: 20 mM, competitive inhibitor
dipicolinic acid
-
-
dipicolinic acid di-imidate
-
-
dipicolinic acid N-oxide
-
0.8 mM 50% inhibition
DL-diaminopimelic acid
-
slightly inhibitory
DL-diaminopimelic acid
-
-
DL-diaminopimelic acid
-
1.2 mM, 91% inhibition
glyoxylic acid
-
-
Isoleucine
-
5 mM, significant inhibition
Isoleucine
-
not inhibitory
KCl
-
1 M, 35% inactivation, 2 M, 55% inactivation
L-alpha-(2-aminoethoxyvinyl)-glycine
-
1.2 mM, 100% inhibition
L-arginine
-
slightly inhibitory
L-arginine
-
-
L-arginine
-
5 mM, significant inhibition
L-arginine
-
-
L-arginine
-
1 mM, 69% inhibition
L-arginine
-
1.2 mM, 62% inhibition
L-aspartate
P19808
competitive inhibition, L-aspartate 4-semialdehyde as varied substrate
L-aspartate 4-semialdehyde
P19808
uncompetitive inhibition by high concentrations of, no overcome by increasing pyruvate concentrations from 5 to 15 mM
L-aspartate 4-semialdehyde
Q9JZR4
is subject to substrate inhibition by high concentrations
L-aspartate 4-semialdehyde
-
DHDPS is subject to substrate inhibition by L-aspartate 4-semialdehyde
L-aspartate 4-semialdehyde
-
competitive inhibition at high substrate concentrations
L-homoserine
-
1 mM, 13% inhibition
L-lysine
-
0.1 mM, complete inhibition, I0.5: 0.02 mM
L-lysine
-
-
L-lysine
-
no inhibition
L-lysine
-
Ki: 0.21 mM
L-lysine
-
no inhibition
L-lysine
-
-
L-lysine
-
1 mM, 50% inhibition
L-lysine
-
no inhibition
L-lysine
-
discussion of plant inhibition versus bacterial inhibition
L-lysine
P19808
lack of feedback inhibition, not regulated under normal physiological conditions
L-lysine
Q5HG25
poor feedback inhibition by
L-lysine
Q9JZR4
is significantly more sensitive to feedback inhibition than Escherichia coli DHDPS
L-lysine
-
at 1 mM 23% residual activity, at 100 mM 13% residual activity
L-lysine
-
no difference in its sensitivity or behaviour with respect to L-lysine when compared to the wild-type
L-lysine
-
binding interaction of L-lysine is characterised as a cooperative event in which an entropic pre-organisation step precedes a secondary enthalpic association. This allosteric association is of a mixed competitive nature in which heterotropic ligand cooperativity is observed to subtly influence the binding events
L-lysine
-
inhibition of the tetrameric wild-type enzyme, but not of the disrupted minimeric mutant enzyme. Allosteric binding by two molecules of (S)-lysine at the DHDPS tight-dimer interface cleft has been observed to operate via a cooperative mechanism and to result in incomplete partial mixed inhibition, inhibition kinetics, overview
L-lysine
Q9I4W3
binds at three sites to the enzyme, binding structure, overview
L-lysine
-
natural feedback inhibitor
L-lysine
-
feedback inhibition
L-lysine
D7U7T8
feedback inhibition, binding of lysine to the allosteric cleft of the enzyme, cooperative binding, structural mechanism for allosteric inhibition, overview. With respect to L-aspartate-4-semialdehyde, lysine is a noncompetitive inhibitor
L-methionine
-
at 10 or 100 mM 80% residual activity
L-threonine
-
at 100 mM 40% residual activity
L-threonine
-
at 100 mM 23% residual activity, at 100 mM 33% residual activity
Lys
-
strong inhibition
lysine
-
inhibition of wild-type DHDPS by lysine with respect to pyruvate is partial and uncompetitive, and partial non-competitive with respect to L-aspartate 4-semialdehyde. Ethanolamine, n-butylamine, 1-amino-2-propanol, 3-amino-1-propanol, iso-butylamine and Tris-HCl cannot rescue activity
NaBH4
-
NaBH4 reduction of the pyruvyl-Schiff-base intermediate results in enzyme inactivation
oxaloacetate
P19808
non-competitive inhibition, pyruvate as varied substrate
PMSF
-
-
pyridine-2,6-dicarboxylic acid
-
-
pyridine-2,6-dicarboxylic acid
-
20 mM, 75% inhibition; 20 mM, 80% inhibition
pyridine-2,6-dicarboxylic acid
-
20 mM, 67% inhibition
pyruvate
A9CFV4, A9CGZ4, A9CHR2, A9CL94, A9CL97, Q7CU96, Q7D0E8, Q7D313, Q7D3Z9, Q8UGL3
substrate inhibition
S-(2-aminoethyl)-L-cysteine
-
-
S-(2-aminoethyl)-L-cysteine
-
-
S-(2-aminoethyl)-L-cysteine
-
I0.5: 0.4 mM
S-(2-aminoethyl)-L-cysteine
-
-
S-(2-aminoethyl)-L-cysteine
-
4.6 mM, 50% inhibition
Sodium borohydride
-
wild-type DHDPS is inactivated when incubated with pyruvate, whereas incubation with L-aspartate 4-semialdehyde has no effect
Succinate-semialdehyde
-
reversible inhibitor which is competitive with respect to L-aspartate-4-semialdehyde and uncompetitive with respect to pyruvate
Succinic semialdehyde
-
-
Succinic semialdehyde
P19808
uncompetitive inhibition, L-aspartate 4-semialdehyde as varied substrate
threo-4-hydroxy-L-lysine
-
-
threo-4-hydroxy-L-lysine
-
-
trans-(1SS,3R5S)-3,5-thiomorpholinedicarboxylic acid, dimethyl ester, 1-oxide
-
20 mM, 20% inhibition
trans-(1SS,3R5S)-3,5-thiomorpholinedicarboxylic acid, dimethyl ester, 1-oxide
-
20 mM, 14% inhibition; 20 mM, 22% inhibition
trans-(1SS,3R5S)-3,5-thiomorpholinedicarboxylic acid, dimethyl ester, 1-oxide
-
20 mM, 12% inhibition
lysine ethyl ester
-
-
additional information
-
not inhibitory: ATP, ADP, AMP, citric acid, L-malic acid, 2-oxoglutaric acid, fumaric acid, succinic acid, L-glutamic acid, L-threonine
-
additional information
-
not inhibitory: iodoacetate, iodoacetamide, p-chloromercuribenzoate
-
additional information
-
not inhibitory: L-Thr, L-Met
-
additional information
-
no inhibition by (S)-aspartate 4-semialdehyde, but inhibition occurs by a derivative that is built of the compounbd in the enzyme preparation by ozonolysis
-
additional information
-
no inhibition by dipicolinic acid methyl ester, (3R,5S)-thiomorpholine-3,5-dicarboxylic acid compound 20b, (3R,5R)-thiomorpholine-3,5-dicarboxylic acid, compound 23, compound 24b, dimethyl (3R,5S)thiomorpholine-3,5-dicarboxylate 1,1-dioxide
-
additional information
Q9X1K9
not inhibited by (S)-lysine, suggesting that feedback control of the lysine biosynthetic pathway evolves later in the bacterial lineage
-
additional information
-
inhibition studies by using 4-oxo-heptenedioic acid analogues, determination of second-order rate constants of inactivation, substrate co-incubation studies show that the inhibitors act at the active-site, interaction analyzed by mass spectrometry, sites of enzyme alkylation determined
-
additional information
-
new constrained inhibitors of DHDPS identified and tested, time-dependent inhibition and substrate protection, dimethyl 2,2'-benzene-1,3-diylbis[(hydroxyimino)ethanoate] discovered as a relatively potent inhibitor of DHDPS enzyme, validates constrained acyclic-intermediate model as a potential inhibitor lead, modifications of the aromatic ring are possible and may result in improvements in activity
-
additional information
-
insensitivity to lysine inhibition
-
additional information
-
no substrate inhibition by (S)-aspartate 4-semialdehyde
-
additional information
-
is not inhibited by 1-100 mM L-lysine or L-isoleucine
-
additional information
-
1-100 mM L-isoleucine has no effect
-
additional information
-
is insensitive to L-lysine inhibition, produces no binding isotherm upon L-lysine addition in either the absence or presence of pyruvate
-
additional information
-
the substrate specificity of the enzyme, two pyruvate analogues, previously classified as weak competitive inhibitors, are turned over productively by DHDPS, NMR spectroscopy, overview
-
additional information
-
no feedback inhibition by L-lysine
-
additional information
-
1,3-phenylene bis(ketoacid) derivatives as enzyme inhibitors, overview. Ketoacid derivatives act as slow and slow-tight binding inhibitors with either an encounter complex or a condensation product for the slow and slow-tight binding inhibitors, respectively, modeling, overview. No or poor inhibition by dimethyl 2,2'-benzene-1,3-diylbis(oxoacetate), (2E,2'E)-2,2'-benzene-1,3-diylbis[(hydroxyimino)ethanoic acid], and dimethyl-2,2'-(2-methoxy-1,3-phenylene)bis(2-oxoacetate)
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
(2R,3S,6S)-2,6-diamino-3-hydroxyheptanedioic acid
-
134% of the activity without
(2R,6S)-2,6-diamino-2-methyl-3-heptenedioic acid
-
110% of the activity without
(2R,6S/S)-2,6-diamino-6-phosphonohexanoic acid
-
113% of the activity without
2-mercaptoethanol
-
activation
cysteine
-
activation
isocitric acid
-
activation
L-lysine
-
mutagenesis of the lysine binding sites of the Corynebacterium glutamicum enzyme according to the residues in the Escherichia coli enzyme does not conver the expected feedback inhibition but an activation of the nezyme by L-lysine
L-methionine
-
at 10 mM 112% activity, at 100 mM 111% activity
meso-diaminopimelate
-
at 1 mM 108% activity, at 10 mM 122% activity
additional information
-
meso-diaminopimelate has no effect
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.21
(R,S)-aspartate 4-semialdehyde
-
pH 8.0, 30C, with pyruvate
-
0.09
(S)-aspartate 4-semialdehyde
-
mutant T44V, pH 8.0, 30C
0.11
(S)-aspartate 4-semialdehyde
-
pH 8.0, 30C, with pyruvate
0.11
(S)-aspartate 4-semialdehyde
-
wild-type enzyme, pH 8.0, 30C
0.11
(S)-aspartate 4-semialdehyde
-
30C
0.11
(S)-aspartate 4-semialdehyde
-
wild-type
0.22
(S)-aspartate 4-semialdehyde
-
similar to those of DHDPS enzyme of Escherichia coli
0.33
(S)-aspartate 4-semialdehyde
Q5HG25
-
0.38
(S)-aspartate 4-semialdehyde
Q8RBI5
by adding 20 mM pyruvate, at 60C
0.39
(S)-aspartate 4-semialdehyde
-
Y107W mutant
0.58
(S)-aspartate 4-semialdehyde
-
mutant Y107F, pH 8.0, 30C
0.58
(S)-aspartate 4-semialdehyde
-
Y107F mutant
0.92
(S)-aspartate 4-semialdehyde
Q8RBI5
modeled with substrate inhibition, at 60C
2.7
(S)-aspartate 4-semialdehyde
-
mutant Y133F, pH 8.0, 30C
0.137
(S)-aspartate-4-semialdehyde
-
recombinant mutant H56K, pH 8.0, temperature not specified in the publication
0.16 - 0.32
(S)-aspartate-4-semialdehyde
A9CFV4, A9CGZ4, A9CHR2, A9CL94, A9CL97, Q7CU96, Q7D0E8, Q7D313, Q7D3Z9, Q8UGL3
pH and temperature not specified in the publication
0.248
(S)-aspartate-4-semialdehyde
-
recombinant mutant E84T, pH 8.0, temperature not specified in the publication
0.3
(S)-aspartate-4-semialdehyde
-
recombinant wild-type enzyme, pH 8.0, temperature not specified in the publication
0.55
DL-aspartate-4-semialdehyde
-
-
0.05
L-aspartate 4-semialdehyde
-
mutant L170E/G191E, at 30C
0.052
L-aspartate 4-semialdehyde
Q9JZR4
at 30C, in 100 mM HEPES buffer, pH 8.0
0.11
L-aspartate 4-semialdehyde
-
at 30C, in 100 mM HEPES buffer, pH 8.0
0.12
L-aspartate 4-semialdehyde
-
mutant K161R, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR; wild-type, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR
0.13
L-aspartate 4-semialdehyde
-
wild-type, at 30C
0.13
L-aspartate 4-semialdehyde
-
mutant T44S
0.14
L-aspartate 4-semialdehyde
-
pH 8.0
0.15
L-aspartate 4-semialdehyde
-
mutants Q196D, D193A and D193Y, at 30C
0.15
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp237
0.17
L-aspartate 4-semialdehyde
-
mutant D193A, at 30C
0.17
L-aspartate 4-semialdehyde
Q9I4W3
pH 8.0, 37C, recombinant His-tagged enzyme
0.18
L-aspartate 4-semialdehyde
-
-
0.18
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp237
0.23
L-aspartate 4-semialdehyde
-
C-terminal truncated DHDPS (H225*), at 30C, in 150 mM HEPES, pH 8.0, 0.16 mM NADPH, 50 microg/ml DHDPR
0.23
L-aspartate 4-semialdehyde
-
mutant K161A, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR
0.23
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp171/Arg237; pH not specified in the publication, 30C, recombinant wild-type enzyme
0.25
L-aspartate 4-semialdehyde
-
wild-type, at 30C
0.29
L-aspartate 4-semialdehyde
-
polyhistidine-tagged wild-type DHDPSR
0.32
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp171/Arg237
0.36
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 45C, recombinant wild-type enzyme
0.42
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp171
0.43
L-aspartate 4-semialdehyde
-
pH 8.0, 30C, recombinant wild-type enzyme
0.46
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp171
0.63
L-aspartate 4-semialdehyde
P19808
reaction mixture includes 100 mM Tris-HCl, 2.5 mM pyruvate, 0.05-0.1 microg dihydrodipicolinate synthase and 0.2 mM NADPH, from 5.6 to 22.4 mM plots deviate from typical Michaelis-Menten kinetics, increased concentration fits an uncompetitive substrate inhibition model
1.1
L-aspartate 4-semialdehyde
-
pH 8.0, 30C, recombinant mutant A204R
1.1
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp168/Asp171
1.2
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp168/Asp237
1.3
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp168/Asp171
1.5
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp168
1.7
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp168/Asp237
2.2
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp168
5.1
L-aspartate 4-semialdehyde
-
mutant enzyme R138A
37
L-aspartate 4-semialdehyde
-
mutant enzyme R138H
0.25
L-aspartate-4-semialdehyde
-
-
0.4
L-aspartate-4-semialdehyde
-
-
0.46
L-aspartate-4-semialdehyde
-
-
0.6
L-aspartate-4-semialdehyde
-
-
0.765
L-aspartate-4-semialdehyde
-
-
0.8
L-aspartate-4-semialdehyde
-
-
1
L-aspartate-4-semialdehyde
-
-
1
L-aspartate-4-semialdehyde
-
-
3.13
L-aspartate-4-semialdehyde
-
-
0.05
pyruvate
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp237
0.07
pyruvate
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp171
0.08
pyruvate
-
mutant T44V, pH 8.0, 30C
0.08
pyruvate
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp171/Arg237; pH not specified in the publication, 30C, recombinant wild-type enzyme
0.1
pyruvate
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp237
0.11
pyruvate
-
similar to those of DHDPS enzyme of Escherichia coli
0.12
pyruvate
Q5HG25
-
0.13
pyruvate
-
-
0.15
pyruvate
-
wild-type, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR
0.15
pyruvate
-
pH not specified in the publication, 45C, recombinant wild-type enzyme
0.16
pyruvate
-
mutant Y107F, pH 8.0, 30C
0.16
pyruvate
-
Y107F mutant
0.16
pyruvate
-
wild-type, at 30C
0.16
pyruvate
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp171
0.17
pyruvate
-
pH 8.0, 30C, recombinant wild-type enzyme
0.18
pyruvate
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp171/Arg237
0.25
pyruvate
-
pH 8.0, 30C, with (S)-aspartate 4-semialdehyde
0.26
pyruvate
-
wild-type enzyme, pH 8.0, 30C
0.26
pyruvate
-
30C
0.26
pyruvate
-
wild-type
0.26
pyruvate
-
at 30C, in 100 mM HEPES buffer, pH 8.0
0.28
pyruvate
-
mutant enzyme R138H
0.3
pyruvate
-
pH 8.0, 30C, with (R,S)-aspartate 4-semialdehyde
0.31 - 0.44
pyruvate
A9CFV4, A9CGZ4, A9CHR2, A9CL94, A9CL97, Q7CU96, Q7D0E8, Q7D313, Q7D3Z9, Q8UGL3
pH and temperature not specified in the publication
0.32
pyruvate
P19808
2.5 mM pyruvate replaced by 0.1-2 mM, no substrate inhibition observed at high concentrations of pyruvate
0.32
pyruvate
-
mutant Q196D, at 30C
0.33
pyruvate
-
pH 8.0, 30C, recombinant mutant A204R
0.36
pyruvate
-
polyhistidine-tagged wild-type DHDPS
0.37
pyruvate
-
C-terminal truncated DHDPS (H225*), at 30C, in 150 mM HEPES, pH 8.0, 0.16 mM NADPH, 50 microg/ml DHDPR
0.38
pyruvate
-
pH 8.0
0.43
pyruvate
-
-
0.44
pyruvate
-
mutant D193A, at 30C
0.45
pyruvate
-
mutant enzyme R138A
0.45
pyruvate
-
mutant K161A, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR
0.46
pyruvate
-
mutant Q234D, at 30C
0.5
pyruvate
-
-
0.5
pyruvate
Q9JZR4
at 30C, in 100 mM HEPES buffer, pH 8.0
0.556
pyruvate
-
recombinant mutant E84T, pH 8.0, temperature not specified in the publication
0.57
pyruvate
-
-
0.57
pyruvate
-
mutant D193Y, at 30C
0.57
pyruvate
-
mutant K161R, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR
0.64
pyruvate
-
recombinant mutant H56K, pH 8.0, temperature not specified in the publication
0.827
pyruvate
-
recombinant wild-type enzyme, pH 8.0, temperature not specified in the publication
0.85
pyruvate
Q8RBI5
by adding 2 mM (S)-aspartate 4-semialdehyde, at 60C
0.9
pyruvate
Q9I4W3
pH 8.0, 37C, recombinant His-tagged enzyme
0.92
pyruvate
-
mutant T44S
1.07
pyruvate
-
-
1.2
pyruvate
-
wild-type, at 30C
1.5
pyruvate
-
Y107W mutant
1.5
pyruvate
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp168/Asp171
1.7
pyruvate
-
-
1.7
pyruvate
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp168; pH not specified in the publication, 30C, recombinant mutant DELTAAsp168/Asp237
2.1
pyruvate
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp168
2.4
pyruvate
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp168/Asp171
2.5 - 3
pyruvate
-
-
2.6
pyruvate
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp168/Asp237
2.83
pyruvate
-
pH and temperature not specified in the publication
3.7
pyruvate
-
mutant L170E/G191E, at 30C
11.7
pyruvate
-
-
35
pyruvate
-
mutant Y133F, pH 8.0, 30C
5
L-aspartate-4-semialdehyde
-
-
additional information
additional information
-
kinetic mechanism
-
additional information
additional information
-
kinetic study: the wild-type enzyme shows a ping pong mechanism, while the monomeric mutant L197D/Y107W shows ternary-complex mechanism
-
additional information
additional information
-
Michaelis-Menten kinetics for wild-type and mutant enzymes, overview
-
additional information
additional information
-
kinetic modeling
-
additional information
additional information
D7U7T8
Michaelis-Menten kinetics
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.12
(S)-aspartate 4-semialdehyde
-
mutant T44V, pH 8.0, 30C
0.7
(S)-aspartate 4-semialdehyde
-
mutant Y133F, pH 8.0, 30C
6.5
(S)-aspartate 4-semialdehyde
-
Y107W mutant
10.8
(S)-aspartate 4-semialdehyde
-
mutant Y107F, pH 8.0, 30C
10.8
(S)-aspartate 4-semialdehyde
-
Y107F mutant
124
(S)-aspartate 4-semialdehyde
-
wild-type enzyme, pH 8.0, 30C
124
(S)-aspartate 4-semialdehyde
-
30C
124
(S)-aspartate 4-semialdehyde
-
wild-type
0.038
L-aspartate 4-semialdehyde
-
mutant enzyme R138H
0.149
L-aspartate 4-semialdehyde
-
mutant enzyme R138A
6.5
L-aspartate 4-semialdehyde
-
mutant D193Y, at 30C
9.8
L-aspartate 4-semialdehyde
-
pH 8.0, 30C, recombinant wild-type enzyme
12.4
L-aspartate 4-semialdehyde
-
mutant D193A, at 30C
35
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp168
39
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp237
52
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp168/Asp171
63
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp171/Arg237
68
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp168/Asp237
88
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp237
97
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp171
108
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp168/Asp171
111
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp168
119
L-aspartate 4-semialdehyde
-
pH 8.0, 30C, recombinant mutant A204R
134
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp171/Arg237
136
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 30C, recombinant wild-type enzyme
138
L-aspartate 4-semialdehyde
-
pH 8.0, 30C, recombinant wild-type enzyme
141
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp168/Asp237
213
L-aspartate 4-semialdehyde
P19808
-
233
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp171
465
L-aspartate 4-semialdehyde
-
pH not specified in the publication, 45C, recombinant wild-type enzyme
223
L-aspartate-4-semialdehyde
-
-
0.038
pyruvate
-
mutant enzyme R138H
0.06
pyruvate
-
mutant K161A, at 30C, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR
0.149
pyruvate
-
mutant enzyme R138A
0.16
pyruvate
-
mutant K161R, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR
1.2
pyruvate
-
C-terminal truncated DHDPS (H225*), at 30C, in 150 mM HEPES, pH 8.0, 0.16 mM NADPH, 50 microg/ml DHDPR
1.7
pyruvate
-
mutant L170E/G191E, at 30C
7.2
pyruvate
-
mutant D193Y, at 30C
9.15
pyruvate
-
mutant D193A, at 30C
9.8
pyruvate
-
pH 8.0, 30C, recombinant wild-type enzyme
10.4
pyruvate
-
mutant T44S
11.8
pyruvate
-
mutant Q234D, at 30C
13.28
pyruvate
-
mutant Q196D, at 30C
35
pyruvate
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp168
39
pyruvate
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp237
45
pyruvate
-
wild-type, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR
46.7
pyruvate
Q9JZR4
at 30C, in 100 mM HEPES buffer, pH 8.0
52
pyruvate
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp168/Asp171
63
pyruvate
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp171/Arg237
68
pyruvate
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp168/Asp237
70
pyruvate
-
ping-pong kinetic mechanism
75
pyruvate
-
polyhistidine-tagged wild-type DHDPS
76
pyruvate
-
-
78
pyruvate
-
wild-type, at 30C
88
pyruvate
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp237
92
pyruvate
-
wild-type, at 30C
97
pyruvate
-
pH not specified in the publication, 30C, recombinant mutant DELTAAsp171
108
pyruvate
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp168/Asp171
111
pyruvate
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp168
119
pyruvate
-
pH 8.0, 30C, recombinant mutant A204R
124
pyruvate
-
30C
124
pyruvate
-
at 30C, in 100 mM HEPES buffer, pH 8.0
134
pyruvate
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp171/Arg237
136
pyruvate
-
pH not specified in the publication, 30C, recombinant wild-type enzyme
138
pyruvate
-
pH 8.0, 30C, recombinant wild-type enzyme
141
pyruvate
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp168/Asp237
233
pyruvate
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp171
465
pyruvate
-
pH not specified in the publication, 45C, recombinant wild-type enzyme
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.26
L-aspartate 4-semialdehyde
-
mutant K161A, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR
573
1.3
L-aspartate 4-semialdehyde
-
mutant K161R, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR
573
34
L-aspartate 4-semialdehyde
-
mutant L170E/G191E, at 30C
573
43
L-aspartate 4-semialdehyde
-
mutant D193Y, at 30C
573
53
L-aspartate 4-semialdehyde
-
mutant D193A, at 30C
573
81
L-aspartate 4-semialdehyde
-
mutant T44S
573
83
L-aspartate 4-semialdehyde
-
mutant D193A, at 30C
573
88
L-aspartate 4-semialdehyde
-
mutant Q196D, at 30C
573
110
L-aspartate 4-semialdehyde
-
pH 8.0, 30C, recombinant mutant A204R
573
320
L-aspartate 4-semialdehyde
-
pH 8.0, 30C, recombinant wild-type enzyme
573
368
L-aspartate 4-semialdehyde
-
wild-type, at 30C
573
380
L-aspartate 4-semialdehyde
-
wild-type, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR
573
600
L-aspartate 4-semialdehyde
-
wild-type, at 30C
573
898
L-aspartate 4-semialdehyde
Q9JZR4
at 30C, in 100 mM HEPES buffer, pH 8.0
573
0.13
pyruvate
-
mutant K161A, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR
31
0.28
pyruvate
-
mutant K161R, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR
31
0.46
pyruvate
-
mutant L170E/G191E, at 30C
31
1.1
pyruvate
-
mutant T44S
31
12.6
pyruvate
-
mutant D193Y, at 30C
31
21
pyruvate
-
mutant D193A, at 30C
31
25.6
pyruvate
-
mutant Q234D, at 30C
31
42
pyruvate
-
mutant Q196D, at 30C
31
76.67
pyruvate
-
wild-type, at 30C
31
93.4
pyruvate
Q9JZR4
at 30C, in 100 mM HEPES buffer, pH 8.0
31
300
pyruvate
-
wild-type, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR
31
360
pyruvate
-
pH 8.0, 30C, recombinant mutant A204R
31
488
pyruvate
-
wild-type, at 30C
31
820
pyruvate
-
pH 8.0, 30C, recombinant wild-type enzyme
31
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
32.4
(2E)-4-oxohept-2-enedioic acid
-
mono-ene inhibitor
1.63
(2E,5E)-4-oxohepta-2,5-dienedioic acid
-
more potent than the corresponding mono-ene inhibitors
0.07
(S)-aspartate 4-semialdehyde
-
substrate inhibition
0.15
(S)-aspartate-4-semialdehyde
A9CFV4, A9CGZ4, A9CHR2, A9CL94, A9CL97, Q7CU96, Q7D0E8, Q7D313, Q7D3Z9, Q8UGL3
pH and temperature not specified in the publication
0.32
(S)-Lys
-
with respect to L-aspartate 4-semialdehyde
3.9
(S)-Lys
-
with respect to pyruvate
0.32
(S)-lysine
-
versus (S)-aspartate 4-semialdehyde
3.9
(S)-lysine
-
versus pyruvate
4.7
(S)-lysine
Q8RBI5
180 mM, at 60C
0.29
2,2'-(2-hydroxy-1,3-phenylene)bis(2-oxoacetic acid)
-
pH 8.0, 30C
-
2.96
2,2'-benzene-1,3-diylbis(oxoacetic acid)
-
49% inhibition at 5 mM, mimics the substrate pyruvate, binding with the active site lysine residue, slow inhibition
2.96
2,2'-benzene-1,3-diylbis(oxoacetic acid)
-
pH 8.0, 30C
0.17
2-oxopimelic acid
-
in 50 mM Tris-HCl (pH 8.2), at 22C
0.21
3-hydroxy-2-oxopropanoate
-
time-dependent inhibition, value similar to that of (S)-lysine
22
4-oxo-1,4-dihydropyridine-2,6-dicarboxylic acid
-
with respect to pyruvate
24.8
4-oxo-1,4-dihydropyridine-2,6-dicarboxylic acid
-
with respect to L-aspartate 4-semialdehyde
10.9
diethyl (2E)-4-oxohept-2-enedioate
-
mono-ene inhibitor
4.95
diethyl (2E,5E)-4-oxohepta-2,5-dienedioate
-
best inhibitor
0.33
dimethyl 2,2'-benzene-1,3-diylbis[(hydroxyimino)ethanoate]
-
15% inhibition at 1 mM, binding with the active site lysine residue, kinetic analysis corresponds to slow-binding model of inhibition
6.9
dimethyl chelidamate
-
with respect to pyruvate
14
dimethyl chelidamate
-
with respect to L-aspartate 4-semialdehyde
0.33
dimethyl-(2E,2'E)-2,2'-benzene-1,3-diylbis[(hydroxyimino)ethanoate]
-
pH 8.0, 30C
0.04
dimethyl-2,2'-(2-hydroxy-1,3-phenylene)bis(2-oxoacetate)
-
pH 8.0, 30C
-
1
dipicolinic acid
-
-
1.2
dipicolinic acid
-
-
0.8
dipicolinic acid N-oxide
-
-
5.4
L-aspartate 4-semialdehyde
-
-
0.049
L-lysine
D7U7T8
versus L-aspartate-4-semialdehyde, pH 8.0, 30C, recombinant enzyme
0.058
L-lysine
Q9JZR4
with (S)-aspartate 4-semialdehyde as substrate
0.18
L-lysine
-
mutant D193A, with respect to (S)-aspartate 4-semialdehyde
0.19
L-lysine
-
mutant Q196D, with respect to (S)-aspartate 4-semialdehyde
0.21
L-lysine
-
-
0.32
L-lysine
-
with (S)-aspartate 4-semialdehyde as substrate
0.4
L-lysine
-
wild-type, with respect to (S)-aspartate 4-semialdehyde
1
L-lysine
-
-
1.7
L-lysine
Q9JZR4
with pyruvate as substrate
2.2
L-lysine
-
mutant D193A, with respect to pyruvate
3.6
L-lysine
-
wild-type, with respect to pyruvate
3.9
L-lysine
-
with pyruvate as substrate
4.1
L-lysine
-
mutant Q196D, with respect to pyruvate
0.12
lysine
-
wild-type, with pyruvate as substrate
0.14
lysine
-
mutant K161A, with pyruvate as substrate; mutant K161R, with L-aspartate 4-semialdehyde as substrate; mutant K161R, with pyruvate as substrate
0.18
lysine
-
wild-type, with L-aspartate 4-semialdehyde as substrate
0.19 - 0.57
pyruvate
A9CFV4, A9CGZ4, A9CHR2, A9CL94, A9CL97, Q7CU96, Q7D0E8, Q7D313, Q7D3Z9, Q8UGL3
pH and temperature not specified in the publication
4.6
S-(2-aminoethyl)-L-cysteine
-
-
0.3
Succinate-semialdehyde
-
in 50 mM Tris-HCl (pH 8.2), at 22C
0.23
lysine
-
mutant K161A, with L-aspartate 4-semialdehyde as substrate
additional information
additional information
-
(S)-lysine allosteric inhibition kinetics and mechanism, overview
-
additional information
additional information
D7U7T8
kinetic and thermodynamics of allosteric inhibitin by lysine, overview
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
20
(2R,6S)-piperidine-2,6-dicarboxylic acid
-
IC50: 20 mM, 83% inhibition at 50 mM
0.065
(S)-lysine
-
pH and temperature not specified in the publication
22
4-oxo-1,4-dihydropyridine-2,6-dicarboxylic acid
-
i.e. chelidamic acid, IC50: 22 mM, 99% inhibition at 50 mM, uncompetitive inhibitor with respect to both substrates
14
dimethyl chelidamate
-
IC50: 14 mM. 99% inhibition at 50 mM, noncompetitive with respect to both substrates
20
dipicolinic acid
-
IC50: 20 mM, competitive inhibitor
0.053
L-lysine
Q9JZR4
-
0.21
L-lysine
-
wild-type DHDPS
0.22
L-lysine
-
C-terminal truncated DHDPS (H225*)
225
L-lysine
Q5HG25
in presence of excess of substrates
660
L-lysine
P19808
5 mM pyruvate, addition of 0-716 mM lysine, lack of feedback inhibition
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.0034
-
purified mutant T44V
0.008
-
purified mutant Y133F
0.27
-
purified mutant Y107F
0.72
-
crude extract of mutant T44S
0.81
-
crude extract
1.11
-
purified enzyme
1.5
-
purified enzyme, recombinant enzyme in the absence of the substrate pyruvate
1.61
-
2fold purified enzyme
1.81
-
purified wild-type enzyme
2
-
crude extract, recombinant enzyme in the absence of the substrate pyruvate
12
-
crude extract, recombinant enzyme in the presence of the substrate pyruvate
14.5
-
purified recombinant mutant L51K, pH 8.0, temperature not specified in the publication
14.52
-
20.7fold purified mutant T44S
19.2
-
purified recombinant mutant A49K, pH 8.0, temperature not specified in the publication
33
-
purified enzyme, recombinant enzyme in the presence of the substrate pyruvate
60.5
-
purified recombinant mutant L51T, pH 8.0, temperature not specified in the publication
81.6
-
purified recombinant mutant A49W, pH 8.0, temperature not specified in the publication
104
-
purified recombinant His-tagged enzyme, pH not specified in the publication, temperature not specified in the publication
174
Q8UGL3
purified recombinant His-tagged enzyme, coupled assay, pH and temperature not specified in the publication
454.2
-
purified recombinant mutant H56K, pH 8.0, temperature not specified in the publication
478.1
-
purified recombinant mutant E84T, pH 8.0, temperature not specified in the publication
500.8
-
purified recombinant wild-type enzyme, pH 8.0, temperature not specified in the publication
559.4
-
purified recombinant mutant A49P, pH 8.0, temperature not specified in the publication
additional information
-
-
additional information
-
-
additional information
-
-
additional information
-
-
additional information
-
-
additional information
-
-
additional information
-
-
additional information
-
attempt to examine the specificity of the active site of DHDPS, co-crystallization with the substrate analogue oxaloacetate, solution of the protein structure indicates that pyruvate rather than oxaloacetic acid bounds in the active site, decarboxylation of oxaloacetate not catalysed by DHDPS, rate of pyruvate production independent of DHDPS concentration, indicating that the decarboxylation of oxaloacetate is occurring by a spontaneous and enzyme-independent mechanism, confirmed by kinetic analysis
additional information
-
archetypal subunit orientation in the crystal structure of other dihydrodipicolinate synthase enzymes not observed, structure refinement will provide information regarding the structural evolution of dihydrodipicolinate synthase and the design of antibiotics targeting lysine biosynthesis in Staphylococcus aureus
additional information
P19808
structure in vicinity of the putative binding site for S-lysine indicates that the allosteric binding site in the Escherichia coli dihydrodipicolinate synthase does not exist in the enzyme of Corynebacterium glutamicum, difference of three non-conservative amino acids substitutions, explains lack of feedback inhibition by lysine in Corynebacterium glutamicum
additional information
-
role of Tyr107 residue in determining the quaternary structure, structural, biophysical, and kinetic studies of the Y107W mutant, catalytic ability and apparent melting temperature reduced by the mutation, tetrameric quaternary structure critical to control specificity, heat stability, and intrinsic activity
additional information
-
titration of (S)-lysine into buffered solutions of MosA protein in the presence or absence of saturating amounts of pyruvate, thermodynamic data for (S)-Lysine binding to MosA protein, non-competitive mechanism with substrate-dependent differences in the energetics of inhibitor binding
additional information
-
substrate protection experiments, inhibition by alkylation the active-site lysine residue (Lys161) and a surface cysteine residue, slower alkylation of other cysteine residues with lower surface accessibility, co-incubation of the enzyme with the inhibitors in the presence of pyruvate protect against active-site alkylation and cysteine residues
additional information
-
analysis of ability of rhizopines to interact with MosA protein in the presence and absence of methyl donors, no methyltransferase activity observed in the presence of scyllo-inosamine and S-adenosylmethionine (SAM), presence of rhizopine compounds does not affect kinetics of dihydrodipicolinate synthesis
additional information
Q8RBI5
specific activity relatively high even at 30C, structural model reveals that the active site is well conserved
additional information
Q5HG25
quaternary structure different from other characterized homologues, dimer both in solution and in the crystal, catalytically important Lys-163 and the proton relay catalytic triad comprising Thr-46, Tyr-109 and Tyr-135 corresponds well with the enzyme homologue of Escherichia coli, deletion mutant lacking the three helical domains reveals a significant reduction in enzymatic activity, changes in the catalytic site upon pyruvate binding provide a structural basis for the ping-pong reaction mechanism, no feedback inhibition by lysine, free and substrate bound forms provide a structural rationale for catalytic mechanism, unique conformational features crucial for the design of specific non-competitive inhibitors
additional information
-
mechanistic insight into catalysis, structural features and the evolution of quarternary structure, MRSA-DHDPS enzyme exists in a monomer-dimer equilibrium in solution, MRSA-DHDPS dimer is catalytically active
additional information
-
role of beta-hydroxypruvate in regulating biosynthesis of dihydrodipicolinate unknown, crystal structure of DHDPS enzyme complexed with beta-hydroxypyruvate solved, active site shows the presence of the inhibitor covalently bound to Lys161, hydroxyl group of inhibitor is hydrogen-bonded to main-chain carbonyl of Ile203, evidence for a catalytic function played by this peptide unit, highly strained torsion angle conserved in active site of other homologous enzyme, points to critical role in catalysis
additional information
-
DHDPS purified in the presence of pyruvate yields a greater amount of recombinant enzyme with 22fold greater specific activity compared with the enzyme purified in the absence of substrate
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.6
-
assay at, measured using a coupled assay with lactate dehydrogenase to detect pyruvate production
7.9 - 8.2
-
-
8
Q8RBI5
specific condensation of pyruvate and (S)-aspartate 4-semialdehyde optimally at
8
Q5HG25
assay at
8
Q9I4W3
assay at
8
-
assay at
8
D7U7T8
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
22
Q9I4W3
assay at
23
-
assay at
25
-
isothermal titration microcalorimetry at
30 - 45
-
assay at 30C and at 45C
30
-
assay at
30
D7U7T8
assay at
37
-
assay at
85
Q8RBI5
specific condensation of pyruvate and (S)-aspartate 4-semialdehyde optimally at, assay performed at 60C, structural model reveals that the active site is well conserved
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30 - 90
Q8RBI5
activity analyzed at
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.24
Q1PDD5
sequence analysis
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
Cucurbita sp.
-
-
Manually annotated by BRENDA team
Q1PDD5
rarely present
Manually annotated by BRENDA team
Medicago truncatula Jemalong 2HA
-
-
-
Manually annotated by BRENDA team
-
isozyme MtDHDPS2 shows highest expression in roots
Manually annotated by BRENDA team
Medicago truncatula Jemalong 2HA
-
isozyme MtDHDPS2 shows highest expression in roots
-
Manually annotated by BRENDA team
Q1PDD5
high level
Manually annotated by BRENDA team
-
highest activities are observed at 16 and 20 days after pollination, followed by a major decline at 24 days after pollination
Manually annotated by BRENDA team
-
isozymes MtDHDPS1, MtDHDPS3, and MtDHDPS4 shows highest expression in mature seeds
Manually annotated by BRENDA team
Medicago truncatula Jemalong 2HA
-
isozymes MtDHDPS1, MtDHDPS3, and MtDHDPS4 shows highest expression in mature seeds
-
Manually annotated by BRENDA team
Phaseolus sp.
-
-
Manually annotated by BRENDA team
Q1PDD5
rarely present
Manually annotated by BRENDA team
Q1PDD5
high level
Manually annotated by BRENDA team
additional information
Q1PDD5
panicle axe
Manually annotated by BRENDA team
additional information
-
organ-specific isozyme expression
Manually annotated by BRENDA team
additional information
Medicago truncatula Jemalong 2HA
-
organ-specific isozyme expression
-
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
Agrobacterium sp. (strain H13-3)
Agrobacterium sp. (strain H13-3)
Agrobacterium sp. (strain H13-3)
Aquifex aeolicus (strain VF5)
Bacillus clausii (strain KSM-K16)
Bartonella henselae (strain ATCC 49882 / Houston 1)
Campylobacter jejuni subsp. jejuni serotype O:2 (strain NCTC 11168)
Campylobacter jejuni subsp. jejuni serotype O:2 (strain NCTC 11168)
Campylobacter jejuni subsp. jejuni serotype O:2 (strain NCTC 11168)
Campylobacter jejuni subsp. jejuni serotype O:2 (strain NCTC 11168)
Campylobacter jejuni subsp. jejuni serotype O:2 (strain NCTC 11168)
Campylobacter jejuni subsp. jejuni serotype O:2 (strain NCTC 11168)
Clostridium botulinum (strain ATCC 19397 / Type A)
Clostridium botulinum (strain Hall / ATCC 3502 / NCTC 13319 / Type A)
Corynebacterium glutamicum (strain ATCC 13032 / DSM 20300 / JCM 1318 / LMG 3730 / NCIMB 10025)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Hahella chejuensis (strain KCTC 2396)
Legionella pneumophila serogroup 1 (strain 2300/99 Alcoy)
Methanocaldococcus jannaschii (strain ATCC 43067 / DSM 2661 / JAL-1 / JCM 10045 / NBRC 100440)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Neisseria meningitidis serogroup B (strain MC58)
Oceanobacillus iheyensis (strain DSM 14371 / JCM 11309 / KCTC 3954 / HTE831)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Rhodopseudomonas palustris (strain ATCC BAA-98 / CGA009)
Rhodopseudomonas palustris (strain ATCC BAA-98 / CGA009)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Staphylococcus aureus (strain COL)
Staphylococcus aureus (strain COL)
Staphylococcus aureus (strain COL)
Staphylococcus aureus (strain MRSA252)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
31270
-
mass spectrometry
649771
31290
-
monomer size of mutant Y107W, deduced from sequence
690914
31310
-
monomer size of wild-type, mass spectrometry
690914
31320
-
monomer size of mutant Y107W, mass spectrometry
690914
31510
Q9JZR4
mass spectrometry
702445
31530
-
purified enzyme, mass spectrometry
701515
31650
-
mass spectrometry
701515
32290
-
mass spectrometry
702448
33780
-
mass spectrometry
701550
34000
Q8RBI5
size of monomer inccluding His-tag, calculated from sequence
692255
36000
Q1PDD5
sequence analysis
706245
64000
-
dimer size of mutant Y107W, analytical ultracentrifugation
690914
73000
P19808
approximating the size of two DHDPS proteins interacting to form a dimer
690628
108000 - 117000
-
gel filtration
33905
112000
-
gel filtration
33912
115000
-
gel filtration
33903
120000
-
gel filtration
677336
123000
-
gel filtration
33902
123000
-
tetramer size of mutant Y107W, analytical ultracentrifugation
690914
124000
-
gel filtration
33906
124000
-
wild-type, sequence analysis
704696
125300
-
tetramer size of wild-type protein, mass spectrometry, mutant form Y107W reveals a mixture of primarily monomer and tetramer in solution
690914
125400
-
tetramer size of mutant Y107W, mass spectrometry
690914
127000
-
gel filtration
33915
130000
-
non-denaturing PAGE
33911
130000
-
gel filtration
714351
133000
Q9X1K9
sedimentation equilibrium analysis
678046
134000
-
calculation from sedimentation and diffusion coefficient, Stokes' radius
33909
150000
Q8RBI5
native protein, gel filtration
692255
158000
P19808
native protein, gel filtration, homotetramer predicted
690628
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
-
x * 31000, SDS-PAGE
?
-
alpha,beta or alpha2beta
?
-
x * 33000, SDS-PAGE
?
Q9FVC8
x * 34680, mass spectrometry
?
Sinorhizobium meliloti L5-30
-
x * 33000, SDS-PAGE
-
dimer
-
2 * 31000, calculated from sequence
dimer
-
2 * 62400, mutant A204R, crystal structure
dimer
Q9I4W3
three-dimensional structure determination shows that DHDPS forms a homodimer which is stabilized by several hydrogen bonds and van der Waals forces at the interface, active site structure, overview. Each monomer is composed of two domains, the N-terminal domain consists of residues 1-224, and forms an 8-fold parallel alpha/beta barrel
homodimer
Q5HG25
in solution and in the crystal, gel filtration, crystallization and dynamic light scattering experiments
homodimer
Staphylococcus aureus COL
-
in solution and in the crystal, gel filtration, crystallization and dynamic light scattering experiments
-
homotetramer
-
-
homotetramer
Q57695
crystallography
homotetramer
Q9JZR4
crystallography
homotetramer
P19808
ribbons rendition of homotetramer in the crystal lattice shown
homotetramer
-
wild-type, mixture of primarily monomer and tetramer in solution determined for mutant form Y107W, hydrodynamic properties of Y107W oligomers calculated by sedimentation velocity analyses
homotetramer
D7U7T8
homotetramer of four identical (beta/alpha)8-barrel monomers, a dimer of tight dimers, with two tight-dimers binding in a head-to-head manner, with the active site situated near the center of the barrel, molecular dynamics simulations
homotetramer
-
the enzyme adopts the canonical homotetrameric structure in both solution and the crystal state
homotetramer
-
the enzyme shows the typical homotetrameric form exhibited by bacterial DHDPS enzymes, present as a dimer of tight dimers. Each monomer consists of an N-terminal domain (residues 1-224) showing a parallel (beta/alpha)8-barrel (TIM barrel) motif. The smaller C-terminal domain, residues 224-292, is comprised of three alpha-helices
tetramer
-
-
tetramer
-
4 * 28000, SDS-PAGE
tetramer
-
4 * 32000-34000, SDS-PAGE
tetramer
-
4 * 37000-38000 + x * 72000, the 72000 Da subunit possibly modifies the structure and kinetic properties, SDS-PAGE
tetramer
-
4 * 32000, Lys161 is the active site, SDS-PAGE
tetramer
-
homotetramer of (alpha/beta)8 barrels, each containing one active site, crystal structure
tetramer
-
dimer of dimers
tetramer
-
crystallography
tetramer
Q2S9K4
crystallography
tetramer
Q9X1K9
4 * 33800, calculated from sequence
tetramer
Q8RBI5
similar to oligomeric states of other homologous enzyme
tetramer
-
4 * 31000, calculated from sequence
tetramer
-
the dimer-dimer interface is small, accounts only 4.3% of the subunit surface area
tetramer
-
4 * 34000, SDS-PAGE, role of quaternary structure in the TIM-barrel family of enzymes, overview. Unlike other DHDPS enzymes, but like many thermophilic enzymes, Tm-DHDPS has a large number of charged residues at the quaternary interface. Removal of electrostatic interactions disrupts quaternary structure
tetramer
-
4 * 62400, crystal structure, the tetrameric structure is not essential for activity in DHDPS from Mycobacterium tuberculosis
tetramer
O67216
the overall quaternary structure of the enzyme consists of four crystallographically independent molecules (A, C, D, and E) forming dimer of dimers. The monomer exhibits a TIM barrel fold and comprises of 11 alpha helices built by 146 amino acids out of which the first 8 helices form the TIM barrel domain and remaining three helices present in the C-terminal. The monomer also has 10 beta strands composed of 42 amino acids. The enzyme molecule has two types of dimer interfaces, weak-dimer interface and tight-dimer interfaces, structure model, overview
trimer
-
3 * 41000, SDS-PAGE
homotetramer
-
crystallography
-
additional information
-
the tetramer-dimer dissociation constant of the enzyme is 3fold tighter in the presence of pyruvate compared with the apo form
additional information
-
disruption of quaternary structure of DHDPS generates a functional monomer that is no longer inhibited by lysine, overview
additional information
Q9I4W3
the recombinant enzyme adopts a characteristic alpha/beta conformation which is retained up to 65C
additional information
-
flexibility and its relationship to quaternary structure and function from molecular dynamic simulations of native tetrameric and mutant dimeric enzyme forms, it shows that the mutant dimeric enzyme form displays high flexibility, resulting in monomer reorientation within the dimer and increased flexibility at the tight-dimer interface, whereas the enzyme tetramer is relatively rigid. The enzyme dimer exhibits disorder within its active site with deformation of critical catalytic residues and removal of key hydrogen bonds that render it inactive
additional information
-
flexibility and its relationship to quaternary structure and function from molecular dynamic simulations show that the native dimeric enzyme form from a methicillin-resistant strain displays high flexibility, resulting in monomer reorientation within the dimer and increased flexibility at the tight-dimer interface and maintains its catalytic geometry and is thus fully functional
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structure analysis, PDB ID 2HMC, structure comparisons; crystal structure analysis, PDB ID 2R8Wm, structure comparisons; crystal structure analysis, PDB ID 3B4U, structure comparisons; crystal structure analysis, PDB IDs 4I7U, 4I7V, and 4I7W, resolution at 1.42-1.69 A, structure comparisons
A9CFV4, A9CGZ4, A9CHR2, A9CL94, A9CL97, Q7CU96, Q7D0E8, Q7D313, Q7D3Z9, Q8UGL3
purified recombinant His-tagged enzyme in the unliganded form and in forms with bound substrate and with bound substrate plus allosteric inhibitor lysine, hanging drop vapour diffusion method, mixing of 0.002 ml of 10 mg/ml protein in 20 mM Tris, pH 8.0, with 0.002 ml of reservoir solution containing 0.1 M Tris, pH 8.5, and 2 M ammonium sulfate, for ligand-bound enzyme from 0.17 M lithium sulfate monohydrate, 0.085 M Tris pH 8.5, 25.5%(w/v) PEG 4000, 15%(v/v) glycerol with addition of 20 mM pyruvate or 20 mM pyruvate and 20 mM lysine, equilibration against 1 ml reservoir solution, 20C, overnight, X-ray diffraction structure determination and analysis at 1.40-1.55 A resolution
Q8UGL3
purified recombinant enzyme, sitting drop vapour diffusion method, mixing of 0.001 ml of 21.8 mg/ml protein in 20 mM Tris-HCl at pH 8.0 and 0.2 M NaCl, with 0.001 ml of reservoir solution containing 4 M sodium phosphate, 0.1 M imidazole, and 0.2 M NaCl at pH 8.0, 18C, 1 week, X-ray diffraction structure determination and analysis at 1.90 A resolution, molecular replacement
O67216
purified recombinant His-tagged enzyme, sitting drop vapour diffusion method, 0.0015 ml of 14.5 mg/ml protein in 20 mM Tris-HCl, pH 8.0 is mixed with 0.0015 ml of reservoir solution containing 20% w/v PEG 6000, 200 mM sodium chloride, 100 mM Tris-HCl, pH 8.0, including 0.02% w/v sodium azide, 20C, method screening, 5 days to 8 weeks, X-ray diffraction structure determination and analysis at 2.5 A resolution
Q9FVC8
in complex with pyruvate, by sitting- and hanging-drop vapor diffusion method, to 2.15 A resolution, shares the same space group, unit cell parameters, and a similar resolution to the structure of substrate unbound DHDPS. Twelve more hydrogen bond interactions at both interfaces in the crystal structure of pyruvate-bound DHDPS relative to the apo structure
-
in the presence of pyruvate, the hanging-drop vapour-diffusion method, at a resolution of 2.15 A. Crystals belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 84.5, b = 124.6, c = 131.0 A, beta = 90.0
-
sitting-drop vapor-diffusion and hanging-drop vapor diffusion
-
crystallized in a number of forms, predominantly using PEG precipitants, estimated solvent content of 41%, best crystal diffracting to beyond 1.9 A resolution
-
in the presence of its substrate pyruvate, by the sitting-drop vapour diffusion method, to 1.2 A resolution. Belongs to space group C2, in contrast to the unbound form, which has trigonal symmetry. Unit-cell parameters are a = 143.4, b = 54.8, c = 94.3 A , beta = 126.3. The crystal volume per protein weight is 2.3 A3 Da-1 (based on the presence of two monomers in the asymmetric unit), with an estimated solvent content of 46%
-
hanging drop method, data collection and refinement statistics, refinement resolution originally extended to 3.0 A, gradually increased to 2.5 A, and finally to 2.2 A, structural similarities to other dihydrodipicolinate synthase
P19808
by the hanging drop-vapour diffusion method, mutants K161A and K161R solved at resolutions of 2.0 and 2.1 A, respectively. They show no changes in their secondary or tertiary structures when compared to the wild-type structure. Crystal structure of mutant K161A with pyruvate bound at the active site solved at a resolution of 2.3 A, reveals a defined binding pocket for pyruvate that is thus not dependent upon lysine 161
-
complexed with pyruvate, the substrate analogs succinate alpha-semialdehyde and alpa-ketopimelic acid, the inhibitor dipicolinic acid, and the natural feedback inhibitor L-lysine, hanging drop vapor diffusion method, in the presence of beta-octyl glucoside using either potassium phosphate buffer (pH 10) or potassium citrate buffer (pH 7.0) as the precipitant
-
crystal structure at 2.5 A resolution
-
crystal structure of mutant enzyme R138H and R138A, hanging-drop vapor diffusion method
-
examination of the specificity of the active site of DHDPS, co-crystallization with the substrate analogue oxaloacetate, data-collection and refinement statistics
-
hanging-drop vapour-diffusion method, crystal struture of native and (S)-lysine-bound dihydropicolinate synthase are presented to 1.9 A and 2.0 A resolution, respectively
-
in complex with beta-hydroxypyruvate, hanging drop vapor-diffusion method, data processing and refinement statistics
-
mutant T44S, crystals are isomorphous to those of the wild-type enzyme, no significant modification in its tertiary or quaternary structure from that of the wild-type enzyme
-
mutant Y107W, hanging-drop vapor diffusion method, diffraction to beyond 2.0 A resolution, data collection and refinement statistics, solid-state structure of the mutant enzyme largely unchanged
-
purified enzyme in complex with pyruvate and substrate analogue succinic acid semialdehyde, hanging drop vapor diffusion method, mixing of 0.003 ml of 8 mg/mL protein in 20 mM Tris-HCl, pH 8.0, with 0.0012 ml of precipitant solution containing 1.8 M K2HPO4, pH 10, and 0.0006 ml of N-octyl-beta-R-glucopyranoside 6% w/v, 4C, 3-4 days, soaking in cryoprotectant solution containing 1.8 M K2HPO4, pH 10, glycerol 20% v/v, and 120 mM succinic acid semialdehyde and 40 mM pyruvate, X-ray diffraction structure determination and analysis at 2.3 A resolution
-
purified recombinant wild-type and mutant enzymes, hanging or sitting drop vapour diffusion method, at 4C and 21C, 0.006 ml protein solutions: about 10 mg/ml protein, 1.8 M K2PO4, pH 10.0, N-octyl-beta-R-glucopyranoside 6% w/v, + 2 ml reservoir solution: 1.8 M K2PO4, pH 10.0, 3-4 days, X-ray diffraction structure determination and analysis at 2.35-2.5 A resolution
-
at 239K using the hanging drop-vapor diffusion method, at 1.5 A resolution. The four subunits of the asymmetric unit assemble to form a tetramer with an approximate 222 symmetry. At the active site, three residues Tyr132, Thr43 and Tyr106 are observed to constitute a catalytic triad. Has a unique extensive dimerdimer interface that is mediated by strong hydrophobic interactions supplemented by two sets of three hydrogen bonds between four polar residues. Belongs to space group P2(1)2(1)2(1) with cell parameters a = 67.03 A, b = 120.52 A, c = 161.1 A
Q2S9K4
purified recombinant enzyme, hanging drop vapour diffusion method, mixing of 0.002 ml of 11 mg/ml protein in 20 mM Tris, pH 7.5, with 0.002 ml of reservoir solution containing 0.2 M magnesium chloride, 10% w/v PEG 8000, 0.1 M Tris chloride, pH 10, at 8C, X-ray diffraction structure determination and analysis at 1.65-1.74 A resolution, molecular replacement
-
by the oil-batch method at 291 K, to 2.2 A resolution. Belongs to space group P21 with unit-cell parameters a = 80.5 A, b = 76.5 A, c = 101.9 A, gamma = 106.9 A. The asymmetric unit contains four DHDPS molecules, forming a homotetramer with approximate 222 symmetry. The overall tertiary structure of DHDPS possesses a (beta/alpha)8-barrel fold (TIM barrel) with three additional alpha-helices (alpha9-alpha11) at the C-terminus of the chain. The beta-strands of the barrel form an intrinsic network of hydrogen-bonding interactions with the neighbouring beta-strands and are oriented in the same directions. The functional residue Lys161, which participates in Schiff-base formation, is located within the beta-barrel and the side chain of Tyr132 sits over this residue
Q57695
hanging-drop vapour-diffusion method, crystallizes in a monoclinic crystal form
-
purified recombinant DHDPS mutant A204R, sitting drop vapour diffusion method at room temperature, 200 nl of 10 mg/ml protein in 20 mM Tris-HCl, 2 mM 2-mercaptoethanol, 250 mM NaCl, 5% v/v glycerol, 10 mM pyruvate, pH 8.0, is mixed with 230 nl reservoir solution containing 2.0 M ammonium sulphate, 100 mM sodium acetate, pH 5.5, X-ray diffraction structure determination and analysis at 2.0 A resolution
-
three-dimensional structure is determined and refined at 2.28 A resolution
P9WP25
to 2.0 A resolution, space group of the crystal is P212121, with unit cell dimensions of a = 80.7, b = 115.7 and c = 132.1 A. The secondary and tertiary structures are remarkably similar to that of Escherichia coli DHDPS. The hydrogen bond lengths within the catalytic triad, and particularly that between Y133 and T44, differ significantly from those of the Escherichia coli enzyme
Q9JZR4
crystal structure at 2.8 A resolution
-
purified recombinant DHDPS, free or in complex with inhibitor (S)-lysine, 15 mg/ml protein in 50 mM Tris-HCl, pH 8.5, at 20C using hanging drop vapour diffusion method, mixing of 0.005 ml protein solution with 0.005 ml well solution containing 30% w/v PEG-3350, 170 mM MgCl2, 70 mM Tris-HCl, pH 8.5, and 6% v/v propylene glycol. Crystals obtained without 6% propylene glycol are soaked in the reservoir containing 20 mg/ml (S)-lysine, X-ray diffraction structure determination and analysis at 2.65-2.85 A resolution
Q9I4W3
purified recombinant His6-tagged enzyme, hanging drop vapour diffusion method, mixing of 0.002 m of 12.5 mg/ml protein solution with 0.002 ml of reservoir solution containing 18% of PEG6000, 0.2 M MgCl2, and 0.1 M TRIS-HCl, pH 7.6, X-ray diffraction structure determination and analysis at 1.6 A resolution, molecular replacement
Q9I4W3
purified recombinant His-tagged enzyme, hanging-drop vapour-diffusion method, 7.6 mg/ml protein in 150 nl solution is mixed with 150 nl of reservoir solution containing 200 mM ammonium sulfate, 100 mM Bis-Tris pH 5.0-6.0, 23-26% w/v PEG 3350, 0.02% w/v sodium azide, X-ray diffraction structure determination and analysis at 2.5 A resolution
-
crystallization conditions are optimized using vapour diffusion in hanging drops at room temperature. Crystal grown in the presence of pyruvate diffracted X-rays to 2.3 A resolution using synchrotron radiation and belonged to the orthorhombic space group C222(1), with unit-cell parameters a = 69.14, b = 138.87, c = 124.13 A
-
structure of MosA protein solved to 1.95 A resolution, data collection and refinement statistics
-
atomic resolution at 1.45 A, crystal structure confirms the dimeric quarternary structure, reveals that the dimerization interface of the MRSA-DHDPS enzyme is more extensive in buried surface area and noncovalent contacts than the equivalent interface in tetrameric DHDPS enzymes from other bacterial species
-
hanging-drop and sitting-drop method, solved in the native form and in complex with pyruvate at 2.3 A and 2.2 A resolution, respectively, single crystal grown in 2 M ammonium sulfate and 0.1 M Bis-Tris pH 6.5 used for data collection, processing and refinement statistics
Q5HG25
X-ray data-collection statistics, best crystal diffracting to beyond 1.45 A resolution
-
by the hanging-drop vapour-diffusion method, to 2.1 A resolution. The crystal belongs to space group P42212, with unit-cell parameters a = b = 105.5, c = 62.4 A, but the R factors remain high following initial processing of the data. The data set is twinned and it is thus reprocessed in space group P2, resulting in a significant reduction in the R factors
-
purified recombinant Tm-DHDPS-DELTAArg-237, vapor diffusion method, mixing of 150 nl protein solution, containing 11.2 mg/ml protein in 20 mM Tris-HCl, pH 8.0, with 150 nl reservoir solution, containing 40% v/v PEG 300, 100 mM phosphate-citrate, buffer, pH 4.2, and 0.02% w/v sodium azide, X-ray diffraction structure determination and analysis at 1.9-2.1 A resolution
-
the X-ray crystal structure is described
Q9X1K9
purifed recombinant detagged enzyme in complex with pyruvate, hanging drop vapour diffusion method, mixing of 0.002 ml of 10 mg/ml protein in 20 mM Tris, 150 mM NaCl, pH 8.0, and 20 mM pyruvate, with 0.002 ml of reservoir solution containing 0.1 M Bis-Tris propane, pH 8.2, 0.2 M sodium bromide, and 20% w/v PEG 3350, equilibration against 1 ml of reservoir solution, 20C, method ooptimization, X-ray diffraction structure determination and analysis at 2.2 A resolution
-
purified recombinant His-tagged enzyme, hanging drop vapor diffusion method, mixing of 0.002 ml of protein in 20 mM pyruvate and 20 mM lysine, with 0.002 ml of reservoir solution containing 0.1 M MES, pH 6.5, 6% v/v PEG 20000, X-ray diffraction structure determination and analysis at 2.40 A resolution
D7U7T8
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.5 - 8
-
pH 6.5, half-life: 5.3 min, pH 8..0, half-life: 20 min
33908
10.5
-
maximal thermal stability at this pH
33908
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
20 - 95
-
purified recombinnat wild-type enzyme
713953
46.5
Q9JZR4
melting temperature in the absence of substrates
702445
50 - 60
-
thermostability is significantly enhanced in the presence of the substrate pyruvate
704696
59.5
-
melting temperature in the absence of substrates
702445
59.8
-
is thermally stabilised by the first substrate, pyruvate, to a greater extent than its Escherichia coli counterpart
702448
60
-
1 min, 50% loss of activity, 5 min, complete inactivation
33902
60
-
inactivation energy: 67300 cal/mol, change in free energy, DELTAF: 22500 cal/mol, change in enthalpy, DELTAH: 66700 cal/mol, change in entropy DELTAS: 133 cal/degree/mol
33908
61.3
-
is thermally stabilised by the first substrate, pyruvate
702448
65
Q9I4W3
the recombinant enzyme adopts a characteristic alpha/beta conformation which is retained up to 65C
715195
74 - 79
-
inactivation energy: 117000 cal/mol, change in free energy, DELTAF: 23400 cal/mol, change in enthalpy, DELTAH: 113500 cal/mol, change in entropy DELTAS: 259 cal/degre e/mol
33905
75
-
pyruvate increases the half-life from 2.7 to 18.3 min
33908
80
-
no reactivation after heating to 80C, pyruvate increases thermostability
33905
90
Q9X1K9
7 h, 40% loss of activity. When the enzyme is incubated at 90C in 8M urea, 60% of the activity is lost after 90 min
678046
90
O67216
the enzyme is stable up to, high-temperature molecular dynamics simulation of the wild-type and mutant enzymes
729694
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
molecular dynamics simulation to analyze the stability of the enzyme
O67216
the secondary and quaternary structure is significantly stabilized in the presence of pyruvate
-
NaCl required for stability, the amount is inversely proportional to the protein concentration
-
repeated freezing/thawing causes loss of activity
-
unstable at all stages of purification, not stabilized by dithioerythritol, glycerol, KCl, (NH4)2SO4, sucrose, divalent cations
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
pH 8.0, 2 M NaCl
-
-20C, pH 6.5
-
-20C, 20 mM Tris-HCl buffer, pH 8.0
-
-20C, 0.06 M phosphate buffer, pH 7.5, 3 months, no loss of activity
-
-20C, 10 mM triethanolamine buffer, pH 7.5, 10 mM 2-mercaptoethanol, stable for at least 6 months
-
-20C, 20 mM Tris-HCl buffer, pH 8.0
-
-20C, addition of glycerol, several months, no loss of activity
-
-20C, 15% loss of activity after 4 d
-
-20C, 25% loss of activity per week
-
4C, 20 mM bis-Tris propane buffer, pH 8.0, 10 mM pyruvate, 300 mM NaCl, 0.01% NaN3, 3 weeks, 20% loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged enzyme 8fold from Escherichia coli strain BL21 (DE3) by immobilized metal affinity chromatography and gel filtration
Q8UGL3
recombinant His-tagged isozyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography; recombinant His-tagged isozyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography; recombinant His-tagged isozyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography; recombinant His-tagged isozyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography; recombinant His-tagged isozyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography; recombinant His-tagged isozyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography; recombinant His-tagged isozyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography; recombinant His-tagged isozyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography; recombinant His-tagged isozyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography; recombinant His-tagged isozyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography
A9CFV4, A9CGZ4, A9CHR2, A9CL94, A9CL97, Q7CU96, Q7D0E8, Q7D313, Q7D3Z9, Q8UGL3
recombinant enzyme from Escherichia coli strain BL21-CodonPlus (DE3)-RIL by two different steps of anion exchange chromatography, dialysis, hydroxyapatite chromatography, another step of anion exchange chromatography, and gel filtration, to homogeneity
O67216
recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) Star by nickel affinity chromatography, cleavage of the His-tag by TEV protease
Q9FVC8
by gel filtration, recombinant enzyme in the absence of the substrate pyruvate (with a yield of 36.3%) and presence of the substrate pyruvate (with a yield of 98%)
-
by sonication, anion-exchange and hydrophobic interaction liquid chromatography
-
to homogeneity
-
recombinant His-tagged enzyme from Escherichia coli strain XL1-Blue by nickel affinity chromatography
-
by anion-exchange chromatography, hydrophobic interaction and gel filtration
-
gel filtration, SDS-PAGE
-
gel filtration, SDS-PAGE
P19808
by centrifugation and gel filtration
-
enzyme expressed from plasmid in strain XL1-Blue, 69fold
-
gel filtration
-
mutant T44S, no pyruvate added to the crude extract prior to sonication, purified by heat shock and ion-exchange chromatography, 20.7fold with a yield of 123%
-
recombinant enzyme 5.7fold from Escherichia coli strain XL-1 Blue
-
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 Star (DE3) by nickel affinity chromatography and gel filtration
-
recombinant wild-type 5.8fold, recombinant mutant Y107F 17.6fold, recombinant mutant T44V 15.8fold, and recombinant mutant Y133F 18fold
-
wild-type and mutants, by gel filtration
-
wild-type DHDPS, and the coupling enzyme, DHDPR, by ammonium sulphate fractionation
-
by centrifugation and gel filtration
Q2S9K4
recombinant enzyme from Escherichia coli strain BL21(DE3) by anion exchange and hydrophobic interaction chromatography, followed by dialysis
-
recombinant His-tagged isozymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
by gel filtration and ultrafiltration
Q57695
by centrifugation and gel filtration, to homogeneity
Q9JZR4
recombinant His-tagged DHDPS from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
Q9I4W3
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, the proteolytic cleavage by TEV protease does not remove the N-terminal His6-tag efficiently
Q9I4W3
recombinant His-tagged DHDPS 9.7fold from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
gel filtration
Q5HG25
gel filtration, SDS-PAGE
-
by centrifugation, anion-exchange liquid chromatography and gel filtration, 2fold, more than 95% pure
-
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography
D7U7T8
recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography, tag cleavage by thrombin, and gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
gene dapA, sequence comparison with Escherichia coli enzyme, expression of His-tagged enzyme in Escherichia coli strain BL21 (DE3)
Q8UGL3
gene dapA10, sequence comparisons, expression of the His-tagged enzyme isozyme in Escherichia coli strain BL21(DE3); gene dapA1, sequence comparisons, expression of the His-tagged enzyme isozyme in Escherichia coli strain BL21(DE3); gene dapA2, sequence comparisons, expression of the His-tagged isozyme in Escherichia coli strain BL21(DE3); gene dapA3, sequence comparisons, expression of the His-tagged enzyme isozyme in Escherichia coli strain BL21(DE3); gene dapA4, sequence comparisons, expression of the His-tagged enzyme isozyme in Escherichia coli strain BL21(DE3); gene dapA5, sequence comparisons, expression of the His-tagged enzyme isozyme in Escherichia coli strain BL21(DE3); gene dapA6, sequence comparisons, expression of the His-tagged enzyme isozyme in Escherichia coli strain BL21(DE3); gene dapA7, sequence comparisons, expression of the His-tagged enzyme isozyme in Escherichia coli strain BL21(DE3); gene dapA8, sequence comparisons, expression of the His-tagged enzyme isozyme in Escherichia coli strain BL21(DE3); gene dapA9, sequence comparisons, expression of the His-tagged enzyme isozyme in Escherichia coli strain BL21(DE3)
A9CFV4, A9CGZ4, A9CHR2, A9CL94, A9CL97, Q7CU96, Q7D0E8, Q7D313, Q7D3Z9, Q8UGL3
gene dapA or Aq_1143, sequence comparisons, recombinant enzyme expression in Escherichia coli strain BL21-CodonPlus (DE3)-RIL
O67216
orf AT2G45440, recombinant His-tagged enzyme expression in Escherichia coli strain BL21 (DE3) Star
Q9FVC8
dapA gene encoding DHDPS amplified and cloned into the pET11a expression vector, expressed in Escherichia coli BL21-DE3
-
PCR product ligated into pCRBluntII-TOPO and transformed into Escherichia coli One Shot TOP10. The dapA insert then amplified, the product, which now contains NdeI and BamHI restriction-endonuclease sites, ligated into pCR-BluntII-TOPO and transformed into Escherichia coli One Shot TOP10, which enables efficient ligation into an expression vector. DapA and linearized pET-11a ligated with T4 DNA ligase and transformed into Escherichia coli BL21(DE3)
-
plasmid pB3935 containing a His-tagged copy of the dapA2 gene and a gene conferring kanamycin resistance, transformed into Escherichia coli BL21(DE3) for protein overexpression
-
expressed in Escherichia coli BL21(DE3), pET24d vector
Q8RBI5
gene dapA, expression of His-tagged enzyme in Escherichia coli strain XL1-Blue
-
expressed in Escherichia coli, BL21 (DE3) strain carrying the Cbot-DHDPS gene (DapA)
-
from pETSA1, overexpressed
-
inserted into the vector pET11a and transformed into Escherichia coli strain CodonPlus BL21 (DE3)
-
expressed in Escherichia coli STBL2, pET23b vector
P19808
gene dapA, expression of wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3)
-
as His-tagged constructs
-
DHDPS mutants expressed in Escherichia coli strain AT997r-
-
DHDPS overexpressed in Escherichia coli AT997recA-, transformed with site-directed mutants based on the pBluescript plasmid pJG001
-
expressed in Escherichia coli XL-1 Blue harbouring the plasmid pJG001
-
expressed in Escherichia coli XL1-blue, pBluescript vector, recombinant protein
-
expressed in Escherichia coli, pJG001 plasmid for site-directed mutagenesis, mutant Y107W expressed in dapA-negative strain AT997r-
-
expression in Escherichia coli
-
expression in Nicotina tabacum
-
expression in Solanum tuberosum
-
gene dapA, overexpression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 Star (DE3)
-
gene dapA, recombinant expression of lysine-insensitive mutants in Escherichia coli strain MG1655 with a yield improved by 46% compared to the wild-type enzyme
-
mutant T44S expressed in dapA-deficient Escherichia coli AT997r-
-
overexpression of wild-type and mutant enzymes in strain XL1-Blue
-
recombinant enzyme expression in Escherichia coli strain XL-1 Blue using plasmid pJG001
-
the dapA and dapA-H225* genes introduced into the vector pUCX, to transform the auxotrophic Escherichia coli XL1-Blue KanRDELTAdapA cells, under the control of a lac promoter/repressor system. Mutations in dapA introduced into the pET-151/D-TOPO plasmid. C-terminal truncated DHDPS (H225*) expressed in Escherichia coli BL21 Star (DE3)
-
wild-type and mutant cloned into plasmid pET-151/D-TOPO and expressed in Escherichia coli BL21 Star (DE3) competent cells
-
ligated into the expression vector pET30a and transformed into the Escherichia coli strain B834 for overexpression
Q2S9K4
DNA and amino acid sequence determination and analysis, recombinant expression in Escherichia coli strain BL21(DE3)
-
isozymes MtDHDPS, MtDHDPS2, MtDHDPS3, and MtDHDPS4, phylogenetic analysis, quantitative RT-PCR expression analysis, the different isogenes are transcriptionally regulated in an organ-specific manner, subcloning in Escherichia coli strain DH5alpha, expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3), coexpresion of His-tagged isozyme MtDHDPS3 with N-terminally FLAG-tagged enzyme AtDHDPS2 from Arabidopsis thliana in Escherichia coli strain BL21(DE3), expression of isozyme MtDHDPS3 in transgenic Arabidopsis thaliana plants under the control of a constitutive 35S promoter via Agrobacterium tumefaciens transfection resulting in an inhibition of enzyme activity
-
into the pET-21a expression vector and introduced into Escherichia coli Rosetta (DE3) strain
Q57695
dapA gene (Rv2753c) is cloned and expressed in Escherichia coli
P9WP25
expression in Escherichia coli
-
into the expression vector pET151-D to give the plasmid pFH02, to transform chemically super-competent Escherichia coli Top10 cells
Q9JZR4
gene dapA, DNA and amino acid sequence determination and analysis, expression of His-tagged DHDPS in Escherichia coli strain BL21(DE3)
Q9I4W3
gene dapA, expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)
Q9I4W3
gene dapA, expression of His-tagged DHDPS in Escherichia coli strain BL21(DE3)
-
recombinant His-tagged protein is expressed in Escherichia coli BL-21(DE3)
-
expressed in Escherichia coli BL21 (DE3), pET11a expression vector
-
expressed in Escherichia coli BL21(DE3), full length dapA gene (DHDPS-FL) and C-terminal variant (DHDPS D228-295) lacking the three helical domains, pET22b vector
Q5HG25
amplified product cloned into a pCR-Blunt IITOP vector, producing the construct pNS01. DHDPS gene then subcloned into the Escherichia coli expression vector Champion pET101/D-TOPO to produce pNS02. Native recombinant protein overexpressed in Escherichia coli BL21 (DE3)
-
expression of N-terminally His6-tagged enzyme with a thrombin cleavage site in Escherichia coli strain BL21 (DE3)
-
gene dapA, expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
D7U7T8
into pBI121 vector in order to obtain fusion construct of the DHDPS sequence with the reporter gene GUS transiently expressed in the onion epidermal cells by particle gun-mediated bombardment. DHDPS cDNA coding sequence subcloned into the pUC18 plasmid and expressed in Escherichia coli strain AT997, an auxotroph lacking DHDPS activity
Q1PDD5
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
high level expression is present in fast-growing tissue and reproductive tissue. The 5'-regulatory sequence of DHDPS contains a GT-1 box and a (CA)n element, which may play a role in regulating the expression of DHDPS
Q1PDD5
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
C139A
O67216
site-directed in silico mutation
L170E/G191E
-
dimeric mutant of the enzyme, retains only 1.8% of the total catalytic activity of the wild-type tetrameric enzyme
P61A/T63L
-
site-directed mutagenesis, the mutant is not inhibited by L-lysine
P61A/T63L/A65H
-
site-directed mutagenesis, the mutant is not inhibited by L-lysine
T63L
-
site-directed mutagenesis, the mutant is not inhibited by L-lysine
T92E
-
site-directed mutagenesis, the mutant is not inhibited by L-lysine
T96E
-
site-directed mutagenesis, the mutant is not inhibited by L-lysine and shows reduced activity compared to the wild-type enzyme
T96E/K68H
-
site-directed mutagenesis, the mutant is not inhibited by L-lysine and shows reduced activity compared to the wild-type enzyme
T96E/K68H/P61A/T63L
-
site-directed mutagenesis, the mutant is activated by L-lysine 3fold shows reduced activity compared to the wild-type enzyme
T96E/K68H/P61A/T63L/A65H
-
site-directed mutagenesis the mutant is activated by L-lysine 5fold shows reduced activity compared to the wild-type enzyme
T96E/K68H/T63L
-
site-directed mutagenesis, the mutant is activated by L-lysine and shows reduced activity compared to the wild-type enzyme
P61A/T63L
-
site-directed mutagenesis, the mutant is not inhibited by L-lysine
-
T63L
-
site-directed mutagenesis, the mutant is not inhibited by L-lysine
-
T92E
-
site-directed mutagenesis, the mutant is not inhibited by L-lysine
-
A49K
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
A49P
-
site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme and is still sensitive to L-lysine
A49W
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
D193A
-
removal of water mediated hydrogen-bond network
D193Y
-
removal of water mediated hydrogen-bond network and introduction of steric bulk to alter surface topology
E84T
-
site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme and is insensitive to L-lysine
H56K
-
site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme and is insensitive to L-lysine
K161A
-
catalytically active, significant decrease in activity. Is not inactivated when incubated with pyruvate and the reducing agent sodium borohydride. Negligible heat production associated with pyruvate binding to the mutant enzyme, consistent with the lack of Schiff base formation
K161A
-
substantially diminished binding affinity of pyruvate, the surrounding active site scaffold is unable to compensate the entropic penalty associated with ligand localisation in the absence of Schiff-base formation
K161R
-
catalytically active, significant decrease in activity. Is not inactivated when incubated with pyruvate and the reducing agent sodium borohydride. Negligible heat production associated with pyruvate binding to the mutant enzyme, consistent with the lack of Schiff base formation
L197D/Y107W
-
site-directed mutagenesis, the mutant forms a monomer that is catalytically active, but with reduced catalytic efficiency, displaying 8% of the specific activity of the wild-type enzyme. The Michaelis constants for the substrates pyruvate and for (S)-aspartate semialdehyde increase by an order of magnitude. L197D/Y107W is expressed as a folded monomer
L51K
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
Q196D
-
removal of hydrogen bonds and charge-charge repulsion, shortened side chain places charged carboxyl groups proximal at interface
Q234D
-
removal of hydrogen bond and charge-charge repulsion with negatively charged E175. Quaternary structure appears closest to that of the wild-type enzyme
R138A
-
activity is approximately 0.1% of wild-type activity, Km-value for L-aspartate 4-semialdehyde is significantly higher than the wild-type value, shows the same IC50 values as the wild-type enzyme, but different partial inhibition patterns
R138H
-
activity is approximately 0.1% of wild-type activity, Km-value for L-aspartate 4-semialdehyde is significantly higher than the wild-type value, shows the same IC50 values as the wild-type enzyme, but different partial inhibition patterns
T44S
-
the active site is intact, returns much but not all activity likely due to the flexibility of Ser44. Increased flexibility in the active site, which appears to facilitate the binding/reaction of substrate analogues
T44V
-
site-directed mutagenesis, reduced activity, structure is similar to the wild-type enzyme
Y107F
-
site-directed mutagenesis, reduced activity, structure is similar to the wild-type enzyme
Y107F
-
mutant, site-directed mutagenesis
Y107W
-
mutant, site-directed mutagenesis
Y133F
-
site-directed mutagenesis, reduced activity, structure is similar to the wild-type enzyme
A49P
Escherichia coli MG1655
-
site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme and is still sensitive to L-lysine
-
E84T
Escherichia coli MG1655
-
site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme and is insensitive to L-lysine
-
H56K
Escherichia coli MG1655
-
site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme and is insensitive to L-lysine
-
K68H
-
site-directed mutagenesis, the mutant is not inhibited by L-lysine
additional information
-
mutagenesis of the lysine binding sites of the Corynebacterium glutamicum enzyme according to the residues in the Escherichia coli enzyme does not conver the expected feedback inhibition but an activation of the nezyme by L-lysine
K68H
-
site-directed mutagenesis, the mutant is not inhibited by L-lysine
-
additional information
-
mutagenesis of the lysine binding sites of the Corynebacterium glutamicum enzyme according to the residues in the Escherichia coli enzyme does not conver the expected feedback inhibition but an activation of the nezyme by L-lysine
-
L51T
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme and is sill sensitive to L-lysine
additional information
-
detailed structural properties of the mutant enzymes based on the crystal structure models
additional information
-
C-terminal truncated DHDPS (H225*), exhibits a dramatic reduction in both solubility and stability. Equilibrating mixture of quaternary states. The substrate, pyruvate, and the feedback inhibitor, (S)-lysine, both have a positive effect on thermostability of C-terminal truncated DHDPS (H225*)
additional information
-
disruption of quaternary structure of DHDPS generates a functional monomer that is no longer inhibited by lysine, overview
additional information
-
feedback inhibition of the Escherichia coli enzyme by lysine is successfully alleviated after substitution of the residues around the inhibitor's binding sites with those of the Corynabacterium glutamicum enzyme
L51T
Escherichia coli MG1655
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme and is sill sensitive to L-lysine
-
additional information
Escherichia coli MG1655
-
feedback inhibition of the Escherichia coli enzyme by lysine is successfully alleviated after substitution of the residues around the inhibitor's binding sites with those of the Corynabacterium glutamicum enzyme
-
additional information
-
expression of dapA gene of E coli, insensitive to feedback-inhibition by L-lysine
A204R
-
site-directed mutagenesis, disruption of the native tetramer formation by targeting the dimer-dimer interface. The mutant enzyme is a dimeric protein with an identical fold and active-site structure to the tetrameric wild-type enzyme
additional information
Q9I4W3
mutant construction by gene replacement of gene dapA (PA1010) via the sacB-based strategy
additional information
-
construction of mutants Tm-DHDPS-DELTAAsp168, DELTAAsp171, or DELTAArg237 by mutating charged residues, reduction of the number of salt bridges at one of the two tetramerization interface of the enzyme and its interactions results in variants with altered quaternary structure, e.g. monomeric, as shown by analytical ultracentrifugation, gel filtration liquid chromatography, and small angle X-ray scattering, and X-ray crystallographic studies, overview
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
drug development
-
the enzyme is a bona fide drug target for treatment of the Crown Gall disease on crops caused by Agrobacterium tumefaciens, rational design of pesticide agents
drug development
O67216
the enzyme is an attractive target for rational antibiotic and herbicide design
synthesis
O67216
the enzyme can be commercially exploited for high-yield production of (S)-lysine
drug development
-
DHDPS is a potential antibiotic target
pharmacology
-
enzyme structure guides design of novel therapeutics
biotechnology
-
enzyme is a target for herbicide and anti-microbial action
drug development
-
the enzyme is an attractive target for the design and synthesis of herbicides and antibiotics
medicine
-
development of species-specific inhibitors of DHDPS as potential antibacterials
biotechnology
Escherichia coli XL1-Blue
-
enzyme is a target for herbicide and anti-microbial action
-
industry
Q2S9K4
considering the industrial application of this protein, such as its use for lysine biosynthesis, stable conformation via tight tetramerization interfaces may make this valuable protein to be more useful
agriculture
-
expression of dapA gene of E coli, insensitive to feedback-inhibition by L-lysine
medicine
-
development of species-specific inhibitors of DHDPS as potential antibacterials
drug development
Q9JZR4
the allosteric binding site of DHDPS may be a good starting point for development of an inhibitor specific to Neisseria meningitidis
drug development
-
the allosteric binding site of DHDPS may be a good starting point for development of an inhibitor specific to Neisseria meningitidis
-
medicine
-
development of species-specific inhibitors of DHDPS as potential antibacterials
pharmacology
Q5HG25
enzyme structure analysis for design of novel therapeutics against bacterial pathogen
pharmacology
-
structure of the enzyme guides the design of novel therapeutics against the methicillin-resistant pathogen
pharmacology
Staphylococcus aureus COL
-
enzyme structure analysis for design of novel therapeutics against bacterial pathogen
-
pharmacology
Staphylococcus aureus MRSA252
-
structure of the enzyme guides the design of novel therapeutics against the methicillin-resistant pathogen
-
drug development
Q9I4W3
the intracellular enzyme dihydrodipicolinate synthase a potential drug target because it is essential for the growth of bacteria while it is absent in humans
additional information
Q9I4W3
the enzyme is not an optimal target for drug development against Pseudomonas aeruginosa