Information on EC 4.3.3.7 - 4-hydroxy-tetrahydrodipicolinate synthase

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

EC NUMBER
COMMENTARY
4.3.3.7
-
RECOMMENDED NAME
GeneOntology No.
4-hydroxy-tetrahydrodipicolinate synthase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
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
-
-
-
-
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
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
elimination
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Biosynthesis of secondary metabolites
-
Lysine biosynthesis
-
lysine biosynthesis I
-
lysine biosynthesis II
-
lysine biosynthesis III
-
lysine biosynthesis VI
-
Metabolic pathways
-
Microbial metabolism in diverse environments
-
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 ACCESSION NO.
COMMENTARY
LITERATURE
BA3935 gene product
-
-
DapA
P0A6L2
-
DapA
Q9JZR4
-
-
DapA2
Q81WN7
-
DHDPA synthase
-
-
DHDPS
-
-
-
-
DHDPS
Q81WN7
-
DHDPS
Escherichia coli XL1-Blue
-
-
-
DHDPS
Q9JZR4
-
-
DHDPS
Shewanella benthica ATCC3392
-
-
-
DHDPS
Sinorhizobium meliloti L5-30
Q92R55
;
-
DHDPS
Q5HG25
-
DHDPS
Staphylococcus aureus COL
Q5HG25
-
-
DHDPS
-
-
DHDPS
Q1PDD5
-
dihydro-dipicolinic acid synthase
-
-
dihydrodipicolinate synthase
-
-
dihydrodipicolinate synthase
Q81WN7
-
dihydrodipicolinate synthase
-
-
dihydrodipicolinate synthase
-
-
dihydrodipicolinate synthase
P0A6L2
-
dihydrodipicolinate synthase
-
-
dihydrodipicolinate synthase
-
-
dihydrodipicolinate synthase
Q9JZR4
-
dihydrodipicolinate synthase
Q9JZR4
-
-
dihydrodipicolinate synthase
-
-
dihydrodipicolinate synthase
-
-
-
dihydrodipicolinate synthase
-
-
dihydrodipicolinic acid synthase
-
-
-
-
dihydrodipocolinate synthase
Q1PDD5
-
dihydropicolinate synthetase
-
-
-
-
MosA
Sinorhizobium meliloti L5-30
-
-
-
MosA protein
Q92R55
-
MosA protein
Sinorhizobium meliloti L5-30
Q92R55
;
-
MRSA-DHDPS
Q6GH13
-
MRSA-DHDPS
Staphylococcus aureus MRSA252
Q6GH13
;
-
pyruvate-aspartic semialdehyde condensing enzyme
-
-
-
-
synthase, dihydrodipicolinate
-
-
-
-
VEG81
-
-
-
-
Vegetative protein 81
-
-
-
-
EC 4.2.1.52
P0A6L2
formerly
additional information
-
the enzyme belongs to the TIM-barrel family of enzymes
CAS REGISTRY NUMBER
COMMENTARY
9055-59-8
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
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
gene dapA
-
-
Manually annotated by BRENDA team
strain XL1-Blue expressing the enzyme from a plasmid, 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
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
D1MH64
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
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
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 ACCESSION NO.
COMMENTARY
LITERATURE
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
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
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
-
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
-
the tetrameric structure is not essential for activity in DHDPS from Mycobacterium tuberculosis
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
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
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
-
-
?
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
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
D1MH64, -
-
-
-
?
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
-
-
-
-
?
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
?
-
D1MH64, -
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
?
-
Escherichia coli XL1-Blue
-
no activity with (R)-aspartate 4-semialdehyde
-
?
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
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
-
-
-
?
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
D1MH64, -
-
-
-
?
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
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
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 ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
no effect: Cd2+, Mg2+, Mn2+, Zn2+
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
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)-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
2,2'-benzene-1,3-diylbis(oxoacetic acid)
-
-
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
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
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
-
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
-
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
D1MH64, -
binds at three sites to the enzyme, binding structure, overview
L-lysine
-
natural feedback 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
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
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
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
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-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]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
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
0.55
-
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
D1MH64, -
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
0.32
-
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
0.57
-
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
0.85
-
pyruvate
-, Q8RBI5
by adding 2 mM (S)-aspartate 4-semialdehyde, at 60C
0.9
-
pyruvate
D1MH64, -
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
2.6
-
pyruvate
-
pH not specified in the publication, 45C, recombinant mutant DELTAAsp168/Asp237
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
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
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]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
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
222982
1.3
-
L-aspartate 4-semialdehyde
-
mutant K161R, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR
222982
34
-
L-aspartate 4-semialdehyde
-
mutant L170E/G191E, at 30C
222982
43
-
L-aspartate 4-semialdehyde
-
mutant D193Y, at 30C
222982
53
-
L-aspartate 4-semialdehyde
-
mutant D193A, at 30C
222982
81
-
L-aspartate 4-semialdehyde
-
mutant T44S
222982
83
-
L-aspartate 4-semialdehyde
-
mutant D193A, at 30C
222982
88
-
L-aspartate 4-semialdehyde
-
mutant Q196D, at 30C
222982
110
-
L-aspartate 4-semialdehyde
-
pH 8.0, 30C, recombinant mutant A204R
222982
320
-
L-aspartate 4-semialdehyde
-
pH 8.0, 30C, recombinant wild-type enzyme
222982
368
-
L-aspartate 4-semialdehyde
-
wild-type, at 30C
222982
380
-
L-aspartate 4-semialdehyde
-
wild-type, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR
222982
600
-
L-aspartate 4-semialdehyde
-
wild-type, at 30C
222982
898
-
L-aspartate 4-semialdehyde
-, Q9JZR4
at 30C, in 100 mM HEPES buffer, pH 8.0
222982
0.13
-
pyruvate
-
mutant K161A, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR
16065
0.28
-
pyruvate
-
mutant K161R, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR
16065
0.46
-
pyruvate
-
mutant L170E/G191E, at 30C
16065
1.1
-
pyruvate
-
mutant T44S
16065
12.6
-
pyruvate
-
mutant D193Y, at 30C
16065
21
-
pyruvate
-
mutant D193A, at 30C
16065
25.6
-
pyruvate
-
mutant Q234D, at 30C
16065
42
-
pyruvate
-
mutant Q196D, at 30C
16065
76.67
-
pyruvate
-
wild-type, at 30C
16065
93.4
-
pyruvate
-, Q9JZR4
at 30C, in 100 mM HEPES buffer, pH 8.0
16065
300
-
pyruvate
-
wild-type, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR
16065
360
-
pyruvate
-
pH 8.0, 30C, recombinant mutant A204R
16065
488
-
pyruvate
-
wild-type, at 30C
16065
820
-
pyruvate
-
pH 8.0, 30C, recombinant wild-type enzyme
16065
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
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
0.32
-
(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
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
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
1
-
dipicolinic acid
-
-
1.2
-
dipicolinic acid
-
-
0.8
-
dipicolinic acid N-oxide
-
-
5.4
-
L-aspartate 4-semialdehyde
-
-
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
0.23
-
lysine
-
mutant K161A, with L-aspartate 4-semialdehyde as substrate
4.6
-
S-(2-aminoethyl)-L-cysteine
-
-
0.3
-
Succinate-semialdehyde
-
in 50 mM Tris-HCl (pH 8.2), at 22C
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
20
-
(2R,6S)-piperidine-2,6-dicarboxylic acid
-
IC50: 20 mM, 83% inhibition at 50 mM
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]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
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
14.52
-
-
20.7fold purified mutant T44S
33
-
-
purified enzyme, recombinant enzyme in the presence of the substrate pyruvate
104
-
-
purified recombinant His-tagged enzyme, pH not specified in the publication, 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
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.6
-
-
assay at, measured using a coupled assay with lactate dehydrogenase to detect pyruvate production
7.9
8.2
-
-
8
-
-
assay at
8
-
-, Q8RBI5
specific condensation of pyruvate and (S)-aspartate 4-semialdehyde optimally at
8
-
Q5HG25
assay at
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
23
-
-
assay at
25
-
-
isothermal titration microcalorimetry at
30
45
-
assay at 30C and at 45C
35
-
-
-
37
-
D1MH64, -
-
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
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30
90
-, Q8RBI5
activity analyzed at
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.24
-
Q1PDD5
sequence analysis
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
Cucurbita sp.
-
-
Manually annotated by BRENDA team
Q1PDD5
rarely present
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
Q1PDD5
high level
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
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
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)
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)
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)
Rhizobium meliloti
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
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
31270
-
-
mass spectrometry
31290
-
-
monomer size of mutant Y107W, deduced from sequence
31310
-
-
monomer size of wild-type, mass spectrometry
31320
-
-
monomer size of mutant Y107W, mass spectrometry
31510
-
-, Q9JZR4
mass spectrometry
31530
-
-
purified enzyme, mass spectrometry
31650
-
-
mass spectrometry
32290
-
-
mass spectrometry
33780
-
-
mass spectrometry
34000
-
-, Q8RBI5
size of monomer inccluding His-tag, calculated from sequence
36000
-
Q1PDD5
sequence analysis
64000
-
-
dimer size of mutant Y107W, analytical ultracentrifugation
73000
-
P19808
approximating the size of two DHDPS proteins interacting to form a dimer
108000
117000
-
gel filtration
112000
-
-
gel filtration
115000
-
-
gel filtration
120000
-
-
gel filtration
123000
-
-
gel filtration
123000
-
-
tetramer size of mutant Y107W, analytical ultracentrifugation
124000
-
-
gel filtration
124000
-
-
wild-type, sequence analysis
125300
-
-
tetramer size of wild-type protein, mass spectrometry, mutant form Y107W reveals a mixture of primarily monomer and tetramer in solution
125400
-
-
tetramer size of mutant Y107W, mass spectrometry
127000
-
-
gel filtration
130000
-
-
non-denaturing PAGE
130000
-
-
gel filtration
133000
-
Q9X1K9, -
sedimentation equilibrium analysis
134000
-
-
calculation from sedimentation and diffusion coefficient, Stokes' radius
150000
-
-, Q8RBI5
native protein, gel filtration
158000
-
P19808
native protein, gel filtration, homotetramer predicted
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
alpha,beta or alpha2beta
?
-
x * 31000, SDS-PAGE
?
-
x * 33000, SDS-PAGE
?
Sinorhizobium meliloti L5-30
-
x * 33000, SDS-PAGE
-
dimer
-
2 * 31000, calculated from sequence
dimer
-
2 * 62400, mutant A204R, crystal structure
dimer
D1MH64, -
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
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
-, Q57695
crystallography
homotetramer
-, Q9JZR4
crystallography
tetramer
-
4 * 32000-34000, SDS-PAGE
tetramer
-
4 * 28000, 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
Q9X1K9, -
4 * 33800, calculated from sequence
tetramer
-, Q8RBI5
similar to oligomeric states of other homologous enzyme
tetramer
-
crystallography
tetramer
-
the dimer-dimer interface is small, accounts only 4.3% of the subunit surface area
tetramer
-, Q2S9K4
crystallography
tetramer
-
4 * 31000, calculated from sequence
tetramer
-
-
tetramer
-
4 * 62400, crystal structure, the tetrameric structure is not essential for activity in DHDPS from Mycobacterium tuberculosis
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
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
D1MH64, -
the recombinant enzyme adopts a characteristic alpha/beta conformation which is retained up to 65C
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
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 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
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
-, P63945
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 50mM 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
D1MH64, -
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, -
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.5
8
-
pH 6.5, half-life: 5.3 min, pH 8..0, half-life: 20 min
10.5
-
-
maximal thermal stability at this pH
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
20
95
-
purified recombinnat wild-type enzyme
46.5
-
-, Q9JZR4
melting temperature in the absence of substrates
50
60
-
thermostability is significantly enhanced in the presence of the substrate pyruvate
59.5
-
-
melting temperature in the absence of substrates
59.8
-
-
is thermally stabilised by the first substrate, pyruvate, to a greater extent than its Escherichia coli counterpart
60
-
-
1 min, 50% loss of activity, 5 min, complete inactivation
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
61.3
-
-
is thermally stabilised by the first substrate, pyruvate
65
-
D1MH64, -
the recombinant enzyme adopts a characteristic alpha/beta conformation which is retained up to 65C
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
75
-
-
pyruvate increases the half-life from 2.7 to 18.3 min
80
-
-
no reactivation after heating to 80C, pyruvate increases thermostability
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
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
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 ACCESSION NO.
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 ACCESSION NO.
LITERATURE
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
-
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 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
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
D1MH64, -
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
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
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
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
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)
-
mutant T44S expressed in dapA-deficient Escherichia coli AT997r-
-
overexpression of wild-type and mutant enzymes in strain XL1-Blue
-
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
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
-, P63945
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)
D1MH64, -
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)
-
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 ACCESSION NO.
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 ACCESSION NO.
COMMENTARY
LITERATURE
L170E/G191E
-
dimeric mutant of the enzyme, retains only 1.8% of the total catalytic activity of the wild-type tetrameric 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
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
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
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
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
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
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
Y133F
-
site-directed mutagenesis, reduced activity, structure is similar to the wild-type 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
-
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 ACCESSION NO.
COMMENTARY
LITERATURE
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
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
-
drug development
D1MH64, -
the intracellular enzyme dihydrodipicolinate synthase a potential drug target because it is essential for the growth of bacteria while it is absent in humans
medicine
-
development of species-specific inhibitors of DHDPS as potential antibacterials
pharmacology
-
structure of the enzyme guides the design of novel therapeutics against the methicillin-resistant pathogen
pharmacology
Q5HG25
enzyme structure analysis for design of novel therapeutics against bacterial 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
-