Information on EC 2.7.1.71 - shikimate kinase

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

EC NUMBER
COMMENTARY
2.7.1.71
-
RECOMMENDED NAME
GeneOntology No.
shikimate kinase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ATP + shikimate = ADP + 3-phosphoshikimate
show the reaction diagram
the enzyme binds substrates randomly and in a synergistic fashion
-
ATP + shikimate = ADP + 3-phosphoshikimate
show the reaction diagram
phosphoryl transfer mechanism of shikimate kinase, overview
-
ATP + shikimate = ADP + 3-phosphoshikimate
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
phospho group transfer
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Biosynthesis of secondary metabolites
-
chorismate biosynthesis from 3-dehydroquinate
-
Metabolic pathways
-
Phenylalanine, tyrosine and tryptophan biosynthesis
-
SYSTEMATIC NAME
IUBMB Comments
ATP:shikimate 3-phosphotransferase
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
AroK
-
-
-
-
AroL
-
-
-
-
AtSK1
Q8GY88
-
AtSK2
Q8GY88
-
kinase (phosphorylating), shikimate
-
-
-
-
kinase, shikimate (phosphorylating)
-
-
-
-
OsSK1
Q5NTH4
-
OsSK2
Q5NTH3
-
OsSK3
Q7X7H9
-
shikimate kinase II
-
-
-
-
SKI
-
-
-
-
SKII
-
-
-
-
type I shikimate kinase, aroK-encoded
-
-
CAS REGISTRY NUMBER
COMMENTARY
9031-51-0
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
two isozymes, AtSK1 and AtSK2, AatSK1 is heat-inducible
UniProt
Manually annotated by BRENDA team
gene aroK
-
-
Manually annotated by BRENDA team
shikimate kinase II; strain K12
-
-
Manually annotated by BRENDA team
shikimic acid kinase I
-
-
Manually annotated by BRENDA team
strain K12; two isoenzymes: SK1 and SK2
-
-
Manually annotated by BRENDA team
Escherichia coli DHPYA-T7
gene aroK
-
-
Manually annotated by BRENDA team
Escherichia coli K12
strain K12
-
-
Manually annotated by BRENDA team
Hansenula henricii
-
-
-
Manually annotated by BRENDA team
; strain 26695
Uniprot
Manually annotated by BRENDA team
; strain SS1
-
-
Manually annotated by BRENDA team
strain 26695
Uniprot
Manually annotated by BRENDA team
enzyme aggregate contains 5 activities: EC 1.1.1.25, EC 2.7.1.71, EC 4.2.1.10, EC 4.2.3.4 and EC 2.5.1.19
-
-
Manually annotated by BRENDA team
enzyme aggregate contains 5 activities: EC 1.1.1.25, EC 2.7.1.71, EC 4.2.1.10, EC 4.2.3.4 and EC 2.5.1.19
-
-
Manually annotated by BRENDA team
enzyme aggregate contains 5 activities: EC 1.1.1.25, EC 2.7.1.71, EC 4.2.1.10, EC 4.2.3.4 and EC 2.5.1.19
-
-
Manually annotated by BRENDA team
enzyme aggregate contains 5 activities: EC 1.1.1.25, EC 2.7.1.71, EC 4.2.1.10, EC 4.2.3.4 and EC 2.5.1.19
-
-
Manually annotated by BRENDA team
ARO 1; enzyme aggregate contains 5 activities: EC 1.1.1.25, EC 2.7.1.71, EC 4.2.1.10, EC 4.2.3.4 and 2.5.1.19
-
-
Manually annotated by BRENDA team
enzyme aggregate contains 5 activities: EC 1.1.1.25, EC 2.7.1.71, EC 4.2.1.10, EC 4.2.3.4 and 2.5.1.19
-
-
Manually annotated by BRENDA team
Saccharomycopsis lipolytica
enzyme aggregate contains 5 activities: EC 1.1.1.25, EC 2.7.1.71, EC 4.2.1.10, EC 4.2.3.4 and EC 2.5.1.19
-
-
Manually annotated by BRENDA team
enzyme aggregate contains 5 activities: EC 1.1.1.25, EC 2.7.1.71, EC 4.2.1.10, EC 4.2.3.4 and EC 2.5.1.19
-
-
Manually annotated by BRENDA team
serotype 5b
-
-
Manually annotated by BRENDA team
Shigella flexneri 8401
serotype 5b
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
evolution
-
the enzyme is a member of the nucleoside monophosphate kinases (NMP kinases) family, which show large conformational changes during catalysis
evolution
-, Q8GY88
AtSK2 belongs to the nucleoside monophosphate kinase family
malfunction
-
aroK gene inactivation in DHPYA-T7 leads to high shikimate accumulation, especially when this inactivation is caused by chromosomal deletion
malfunction
-
mutations of the conserved threonine residues associated with the labile C8-H cause the enzymes to lose their saturation kinetics over the concentration range tested
malfunction
Escherichia coli DHPYA-T7
-
aroK gene inactivation in DHPYA-T7 leads to high shikimate accumulation, especially when this inactivation is caused by chromosomal deletion
-
metabolism
-
the enzyme catalyzes the fifth step in the shikimate pathway
metabolism
-
shikimate kinase is the fifth enzyme in the shikimate pathway
metabolism
P56073
shikimate kinase catalyzes the fifth step of the shikimate pathway for biosynthesis of aromatic amino acids
physiological function
-
the Group 2 kinase, shikimate kinase, is controlled by the C8-H of ATP, relationship between the role C8-H of ATP in the reaction mechanism and the ATP concentration as they influence the saturation kinetics of the enzyme activity, regulatory mechanism, overview. The kinase enzyme achieves 2500fold variation in KM through a combination of the various conserved push and pull mechanisms associated with the release of C8-H, the proton transfer cascades unique to the class of kinase in question and the resultant/concomitant creation of a pentavalent species from the gamma-phosphate group of ATP
physiological function
-
he phosphate binding domain in the apo-enzyme is fairly rigid and largely protected from solvent access, even at relatively high temperatures. The shikimate binding domain is highly flexible, the apo-enzyme tends to exhibit large conformational changes to permit LID closure after the shikimate binding. The nucleotide binding domain is initially conformationally rigid, which seems to favour the initial orientation of ADP/ATP, but becomes highly flexible at temperatures above 30C, which may permit domain rotation. Part of the LID domain, including the phosphate binding site, is partially rigid, while another part is highly flexible and accessible to the solvent, mide H/D exchange and mass spectrometry
physiological function
-
shikimate kinase is vital for the survival of Mycobacterium tuberculosis
metabolism
-, Q8GY88
shikimate kinase catalyzes an intermediate step in the shikimate pathway to aromatic amino acid biosynthesis
additional information
-
shikimate binding to the enzyme, docking analysis, conformation of ternary dead-end enzyme-shikimate-ADP complex, molecular dynamics simulation of a fully hydrated model of the docked complex, overview
additional information
-
comparative modeling approach, molecular dynamics calculations using the Escherichia coli structure as template, prediction of in silico and disordered regions, enzyme structure analysis and modeling, overview
additional information
-
modeling of the shikimate-binding pocket with main residues involved in intermolecular interactions with shikimate, overview
additional information
P56073
detailed structure-activity relationship analysis, overview. The critical conserved residues D33, F48, R57, R116, and R132 interact with shikimate. A characteristic three-layer architecture and a conformationally elastic region consisting of F48, R57, R116, and R132 are occupied by shikimate
additional information
-, Q8GY88
mechanism of thermal regulation, computational analysis of AtSK1 and AtSK2 structural variation, overview
additional information
Shigella flexneri 8401
-
comparative modeling approach, molecular dynamics calculations using the Escherichia coli structure as template, prediction of in silico and disordered regions, enzyme structure analysis and modeling, overview
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2'-deoxyATP + shikimate
2'-deoxyADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
P0A4Z2
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-, P56073
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
energy charge plays a role in regulating shikimate kinase, thereby controlling the shikimate pathway
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
SK2 is the isoenzyme that normally functions in aromatic biosynthesis in the cell, SK1 functions only when high intracellular levels of shikimate occurs
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
the enzyme catalyzes the committed step in the seven-step biosynthesis of chorismate
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-, P56073
fifth step in the shikimate pathway
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
the substrate binds in a pocket lined with hydrophobic residues and interacts with several highly conserved charged residues including Asp34, Arg58,Glu61 and Arg136 which project into the cavity
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
Escherichia coli K12
-
-, SK2 is the isoenzyme that normally functions in aromatic biosynthesis in the cell, SK1 functions only when high intracellular levels of shikimate occurs
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
Escherichia coli K12
-
-
-
-
?
ATP + shikimate
ADP + 3-phosphoshikimate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + 3-phosphoshikimate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + 3-phosphoshikimate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + 3-phosphoshikimate
show the reaction diagram
P56073
-
-
-
?
ATP + shikimate
ADP + 3-phosphoshikimate
show the reaction diagram
-, Q8GY88
-
-
-
?
ATP + shikimate
ADP + 3-phosphoshikimate
show the reaction diagram
-
specific phosphorylation of the 3-hydroxy group of shikimate
-
-
?
ATP + shikimate
ADP + 3-phosphoshikimate
show the reaction diagram
Shigella flexneri 8401
-
-
-
-
?
ATP + shikimate
ADP + 3-phosphoshikimate
show the reaction diagram
Escherichia coli DHPYA-T7
-
-
-
-
?
additional information
?
-
-
no activity with XTP, ITP, GTP, TTP, CTP and UTP
-
-
-
additional information
?
-
-, Q5NTH3, Q5NTH4, Q7X7H9
differential expression of the three rice SK genes OsSK1, OsSK2, and OsSK3, and the accompanying changes in the production of shikimate 3-phosphate may contribute to the defense response and to panicle development in rice
-
-
-
additional information
?
-
-, Q5NTH3, Q5NTH4, Q7X7H9
differential expression of the three rice SK genes OsSK1,OsSK2, and OsSK3, and the accompanying changes in the production of shikimate 3-phosphate may contribute to the defense response and to panicle development in rice
-
-
-
additional information
?
-
-, Q8GY88
AtSK2 shikimate binding residues include Gly177, Gly178, Gly179, and Asp130 of the Walker B motif and Phe146, Phe154, Glu158, Arg155, and Arg243
-
-
-
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
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
energy charge plays a role in regulating shikimate kinase, thereby controlling the shikimate pathway
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
SK2 is the isoenzyme that normally functions in aromatic biosynthesis in the cell, SK1 functions only when high intracellular levels of shikimate occurs
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-
the enzyme catalyzes the committed step in the seven-step biosynthesis of chorismate
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
-, P56073
fifth step in the shikimate pathway
-
-
?
ATP + shikimate
ADP + 3-phosphoshikimate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + 3-phosphoshikimate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + 3-phosphoshikimate
show the reaction diagram
-
-
-
-
?
ATP + shikimate
ADP + 3-phosphoshikimate
show the reaction diagram
P56073
-
-
-
?
ATP + shikimate
ADP + 3-phosphoshikimate
show the reaction diagram
-, Q8GY88
-
-
-
?
ATP + shikimate
ADP + 3-phosphoshikimate
show the reaction diagram
Shigella flexneri 8401
-
-
-
-
?
ATP + shikimate
ADP + 3-phosphoshikimate
show the reaction diagram
Escherichia coli DHPYA-T7
-
-
-
-
?
ATP + shikimate
ADP + shikimate 3-phosphate
show the reaction diagram
Escherichia coli K12
-
SK2 is the isoenzyme that normally functions in aromatic biosynthesis in the cell, SK1 functions only when high intracellular levels of shikimate occurs
-
?
additional information
?
-
-, Q5NTH3, Q5NTH4, Q7X7H9
differential expression of the three rice SK genes OsSK1, OsSK2, and OsSK3, and the accompanying changes in the production of shikimate 3-phosphate may contribute to the defense response and to panicle development in rice
-
-
-
additional information
?
-
-, Q5NTH3, Q5NTH4, Q7X7H9
differential expression of the three rice SK genes OsSK1,OsSK2, and OsSK3, and the accompanying changes in the production of shikimate 3-phosphate may contribute to the defense response and to panicle development in rice
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
ATP
-
the adenyl moiety of ATP plays a direct role in the regulation of ATP binding and/or phosphoryl transfer, role of the C8 proton of ATP and conserved Thr residues interacting with the C8-H in the catalysis of shikimate kinase, mechanism, overview
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
requirement for Mg2+ can partially be replaced by Mn2+, Ca2+, Co2+ and Cd2+
Ca2+
-
divalent cation required, Mg2+ and Mn2+ are most effective, Ca2+ activates to a lesser extent
Ca2+
-
divalent cations required, Mg2+ is most effective, but significant activity is obtained with Fe2+, Ca2+, Mn2+ or Co2+, isoenzyme SK2
Cd2+
-
requirement for Mg2+ can partially be replaced by Mn2+, Ca2+, Co2+ and Cd2+
Cl-
-
increases the stability of the structure of the enzyme and influences the affinity of ADP for shikimate kinase
Co2+
-
requirement for Mg2+ can partially be replaced by Mn2+, Ca2+, Co2+ and Cd2+
Co2+
-
divalent cation required, Mg2+ and Mn2+ are most effective, Co2+ activates to a lesser extent
Co2+
-
divalent cations required, Mg2+ is most effective, but significant activity is obtained with Fe2+, Ca2+, Mn2+ or Co2+, isoenzyme SK2
Fe2+
-
divalent cation required, Mg2+ and Mn2+ are most effective, Fe2+ activates to a lesser extent
Fe2+
-
divalent cations required, Mg2+ is most effective, but significant activity is obtained with Fe2+, Ca2+, Mn2+ or Co2+, isoenzyme SK2
Mg2+
-
requirement for Mg2+ can partially be replaced by Mn2+, Ca2+, Co2+ and Cd2+
Mg2+
-
divalent cation required, Mg2+ and Mn2+ are most effective. Optimal Mg2+ concentration is 10 mM
Mg2+
-
divalent cations required, Mg2+ is most effective, but significant activity is obtained with Fe2+, Ca2+, Mn2+ or Co2+, isoenzyme SK2
Mg2+
-
divalent cation required, Mg2+ is most effective followed by Mn2+. Optimal Mg2+ concentration is 2.4 mM
Mg2+
-
in the enzyme that contains bound MgADP- the binding of Mg2+ in the active site involves direct interaction with two protein side-chains
Mg2+
-
influences the position of the hydroxyl groups of the shikimate molecule and some of the residues of the active site of the enzyme
Mg2+
-
required for activity
Mg2+
-
required
Mg2+
-
required
Mg2+
P56073
required
Mg2+
-, Q8GY88
required, Mg2+ coordinating residues are Thr112 and Asp128
Mn2+
-
requirement for Mg2+ can partially be replaced by Mn2+, Ca2+, Co2+ and Cd2+
Mn2+
-
divalent cation required, Mg2+ and Mn2+ are most effective
Mn2+
-
divalent cations required, Mg2+ is most effective, but significant activity is obtained with Fe2+, Ca2+, Mn2+ or Co2+, isoenzyme SK2
Mn2+
-
divalent cation required, Mg2+ is most effective followed by Mn2+
Ni2+
-
divalent cation required, Mg2+ and Mn2+ are most effective, Ca2+ activates to a lesser extent
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
(4R,7S,8aS)-4-[3-(morpholin-4-yl)-3-oxopropyl]-7-[[4-(trifluoromethoxy)benzyl]amino]hexahydropyrrolo[1,2-a]pyrazin-1(2H)-one
-
-
(4R,7S,8aS)-4-[3-oxo-3-(piperidin-1-yl)propyl]-7-[[4-(trifluoromethoxy)benzyl]amino]hexahydropyrrolo[1,2-a]pyrazin-1(2H)-one
-
-
(NH4)2SO4
-
5 mM, 29% inhibition
1-(4-chloro-2,5-dimethoxyphenyl)-1H-tetrazole-5-thiol
-
inhibitor identified by structure-based virtual screening, docking simulations
1-[(3-ethyl-2,6-dimethylquinolin-4-yl)sulfanyl]methanediamine
-
inhibitor identified by structure-based virtual screening, docking simulations
1-[(3S,5S)-5-[3-(1,3-benzodioxol-5-yl)-1,2,4-oxadiazol-5-yl]-1-methylpyrrolidin-3-yl]-3-propan-2-ylurea
-
-
2-(3-methyl-5-sulfanyl-4H-1,2,4-triazol-4-yl)-1-(1,2,3,4-tetrahydro-9H-carbazol-9-yl)ethanone
-
inhibitor identified by structure-based virtual screening, docking simulations
2-(3-methyl-5-sulfanyl-4H-1,2,4-triazol-4-yl)-1-(1,2,3,4-tetrahydro-9H-carbazol-9-yl)ethanone
-
-
2-([[3-([(3R,4S)-4-[2-(4-methylpiperazin-1-yl)-2-oxoethyl]piperidin-3-yl]methyl)-1,2-oxazol-5-yl]methyl]carbamoyl)benzoic acid
-
-
2-[(6-methyl-4-oxo-3-phenyl-3,4,5,6,7,8-hexahydro[1]benzothieno[2,3-d]pyrimidin-2-yl)sulfanyl]acetamide
-
inhibitor identified by structure-based virtual screening, docking simulations
3-methoxy-4-[[2-([2-methoxy-4-[(4-oxo-2-thioxo-1,3-thiazolidin-5-ylidene)methyl]phenoxy]methyl)benzyl]oxy]benzaldehyde
-
noncompetitive inhibitor with respect to both shikimate and MgATP
3-phenyl-4-(5-sulfanyl-1H-tetrazol-1-yl)butanoic acid
-
inhibitor identified by structure-based virtual screening, docking simulations
3-[(8-methoxy-2-methyl-3-propylquinolin-4-yl)sulfanyl]propanoic acid
-
inhibitor identified by structure-based virtual screening, docking simulations
5-bromo-2-(5-[[1-(3,4-dichlorophenyl)-3,5-dioxo-4-pyrazolidinylidene]methyl]-2-furyl)benzoic acid
-
competitive inhibitor toward shikimate and noncompetitive inhibitor with respect to MgATP
5-[(6S)-5-[[5-(hydroxymethyl)furan-2-yl]methyl]-4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridin-6-yl]-3-[4-(trifluoromethoxy)phenyl]-2H-1,2,4-oxadiazol-1-ium
-
-
5-[[(4-fluorobenzyl)sulfanyl]methyl]-4-methyl-4H-1,2,4-triazole-3-thiol
-
inhibitor identified by structure-based virtual screening, docking simulations
6-[3-(1,3-benzodioxol-5-yl)-1,2,4-oxadiazol-5-yl]-5-(3-phenylpropyl)-3a,4,5,6,7,7a-hexahydro-1H-imidazo[4,5-c]pyridine
-
-
ADP
-
4 mM, 44% inhibition
ADP
-
0.5 mM; 12% inhibition
AMP
-
4 mM, 27% inhibition
caffeic acid
-
1 mM, 44% inhibition
N-(3-chloro-4-methylphenyl)-2-[(1-methyl-1H-tetrazol-5-yl)sulfanyl]acetamide
-
inhibitor identified by structure-based virtual screening, docking simulations
Na2SO4
-
5 mM, 27% inhibition
NaCl
-
250 mM, 50% inhibition
NSC162535
P56073
selective inhibitor, identification and binding analysis with enzyme mutant E144A by virtual docking analysis, isothermal titration calorimetry, and crystals structure analysis revealing an induced-fit mechanism, inactivation mechanism, detailed overview. Binding kinetics of wild-type and mutant enzymes
-
p-coumaric acid
-
1 mM, 14% inhibition
shikimate 3-phosphate
-
-
[(4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)sulfanyl]acetic acid
-
inhibitor identified by structure-based virtual screening, docking simulations
[[1,3-dimethyl-9-(2-methylprop-2-en-1-yl)-2,6-dioxo-2,3,6,9-tetrahydro-1H-purin-8-yl]sulfanyl]acetic acid
-
inhibitor identified by structure-based virtual screening, docking simulations
ethyl 4-[([(6S)-6-[4-(propan-2-yl)furan-2-yl]-3,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl]carbonyl)amino]benzoate
-
-
additional information
-
no inhibition by 1 mM Phe, Tyr, Trp, chorismate or prephenate
-
additional information
-
no inhibition by shikimate up to 10 mM
-
additional information
-
screening for the dipeptide inhibitor using in silico structure-based design approach, docking analysis, overview
-
additional information
-
enzyme structure analysis and modeling for rational drug designing, overview
-
additional information
-
molecular docking simulations, Re-docking and cross-docking, and virtual screening for potential inhibitors, analysis of interactions between inhibitors and enzyme residues, overview
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.101
-
ATP
-, P56073
in 100 mM Tris-HCl-KOH buffer, pH 7.5, 50 mM KCl, 5 mM MgCl2, 1.6 mM shikimic acid, 2.5 mM ATP, 1 mM phosphoenolpyruvate, 0.1 mM NADH, 2.5 units of pyruvate kinase/ml, and 2.7 units of lactate dehydrogenase/ml, at 25C
0.101
-
ATP
P56073
enzyme mutant M10A, pH 7.5, 25C; wild-type enzyme, pH 7.5, 25C
0.11
-
ATP
-
pH 9.0
0.143
-
ATP
P56073
enzyme mutant E114A, pH 7.5, 25C
0.16
-
ATP
-
isoenzyme SK2, at 1 mM shikimate
0.231
-
ATP
P56073
enzyme mutant F48Y, pH 7.5, 25C
0.38
-
ATP
-
pH 8.6
0.039
-
shikimate
P56073
enzyme mutant E114A, pH 7.5, 25C
0.06
-
shikimate
-, P56073
in 100 mM Tris-HClKOH buffer, pH 7.5, 50 mM KCl, 5 mM MgCl2, 1.6 mM shikimic acid, 2.5 mM ATP, 1 mM phosphoenolpyruvate, 0.1 mM NADH, 2.5 units of pyruvate kinase/ml, and 2.7 units of lactate dehydrogenase/ml, at 25C
0.06
-
shikimate
P56073
wild-type enzyme, pH 7.5, 25C
0.075
-
shikimate
-
pH 7.0, 25C, mutant enzyme C13S
0.135
-
shikimate
P56073
enzyme mutant M10A, pH 7.5, 25C
0.2
-
shikimate
-
isoenzyme SK2, at 5 mM ATP
0.2
-
shikimate
-
pH 9.0
0.2
-
shikimate
-
-
0.25
-
shikimate
-
pH 8.6
0.28
-
shikimate
-
pH 7.0, 25C, mutant enzyme C162S
0.291
-
shikimate
P56073
enzyme mutant F48Y, pH 7.5, 25C
5
-
shikimate
-
above, isoenzyme SK1, at 5 mM
20
-
shikimate
-
-
0.67
-
ATP
-
pH 7.0, 25C, mutant enzyme C162S
additional information
-
additional information
-
-
-
additional information
-
additional information
-
steady-state kinetics, analysis of wild-type an dmutant enzymes, overview
-
additional information
-
additional information
-
kinetic analysis and conformational dynamics, overview
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
23
-
ATP
-
pH 7.0, 25C, mutant enzyme C13S
35
-
ATP
-
pH 7.0, 25C, wild-type enzyme
40
-
ATP
-
pH 7.0, 25C, mutant enzyme C162S
23
-
shikimate
-
pH 7.0, 25C, mutant enzyme C13S
35
-
shikimate
-
pH 7.0, 25C, wild-type enzyme
40
-
shikimate
-
pH 7.0, 25C, mutant enzyme C162S
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.00948
-
3-methoxy-4-[[2-([2-methoxy-4-[(4-oxo-2-thioxo-1,3-thiazolidin-5-ylidene)methyl]phenoxy]methyl)benzyl]oxy]benzaldehyde
-
in 100 mM TrisHCl (pH 8.0), 50 mM KCl, 5 mM MgCl2, 2 mM ATP, 2 mM phosphoenolpyruvate, 0.7 mM NADH, 3 U/ml proteinase K, 2.5 U/ml lactate dehydrogenase, and 2 mM shikimate, at 25C; pH 8.0, 25C
0.00219
-
5-bromo-2-(5-[[1-(3,4-dichlorophenyl)-3,5-dioxo-4-pyrazolidinylidene]methyl]-2-furyl)benzoic acid
-
in 100 mM TrisHCl (pH 8.0), 50 mM KCl, 5 mM MgCl2, 2 mM ATP, 2 mM phosphoenolpyruvate, 0.7 mM NADH, 3 U/ml proteinase K, 2.5 U/ml lactate dehydrogenase, and 2 mM shikimate, at 25C; pH 8.0, 25C
0.000005
-
asxe1
-
pH and temperature not specified in the publication
-
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0055
-
3-methoxy-4-[[2-([2-methoxy-4-[(4-oxo-2-thioxo-1,3-thiazolidin-5-ylidene)methyl]phenoxy]methyl)benzyl]oxy]benzaldehyde
-
in 100 mM TrisHCl (pH 8.0), 50 mM KCl, 5 mM MgCl2, 2 mM ATP, 2 mM phosphoenolpyruvate, 0.7 mM NADH, 3 U/ml proteinase K, 2.5 U/ml lactate dehydrogenase, and 2 mM shikimate, at 25C
0.0064
-
5-bromo-2-(5-[[1-(3,4-dichlorophenyl)-3,5-dioxo-4-pyrazolidinylidene]methyl]-2-furyl)benzoic acid
-
in 100 mM TrisHCl (pH 8.0), 50 mM KCl, 5 mM MgCl2, 2 mM ATP, 2 mM phosphoenolpyruvate, 0.7 mM NADH, 3 U/ml proteinase K, 2.5 U/ml lactate dehydrogenase, and 2 mM shikimate, at 25C
0.0049
-
NSC162535
P56073
pH 7.5, 25C
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.05718
-
-
-
100
-
-
isoenzyme SK2
100
-
-
shikimate kinase II
101.9
-
-
pH 6.8, 37C
410
-
-
isoenzyme SK1
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.5
-
P56073
assay at
8.6
9
-
-
9
-
-
or higher, in Tris-HCl buffer or bis-Tris propane buffer
9
-
-
assay at
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6
9.5
-
pH 6.0: about 50% of maximal activity, pH 9.5: optimum
6.2
9
-
pH 6.2: about 20% of maximal activity in Tris propane buffer, pH 7.8: about 30% of maximal activity, pH 9.0: optimum
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
P56073
assay at
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.61
-
-
sequence calculation
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-, Q5NTH3, Q5NTH4, Q7X7H9
OsSK1 is induced by treatment with the elicitor N-acetylchitoheptaose; OsSK2 is induced by treatment with the elicitor N-acetylchitoheptaose
Manually annotated by BRENDA team
-, Q5NTH3, Q5NTH4, Q7X7H9
expression of OsSK1 is upregulated specifically during the heading stage of panicle development; expression of OsSK3 is upregulated specifically during the heading stage of panicle development
Manually annotated by BRENDA team
-
isoform SK2 is predominantly expressed early in embryogenesis and vegetative tissues throughout development, isoform SK1 is expressed near or below background levels in vegetative tissues and is only expressed at higher levels in mature embryos and senescing leaves
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
Aquifex aeolicus (strain VF5)
Bacteroides thetaiotaomicron (strain ATCC 29148 / DSM 2079 / NCTC 10582 / E50 / VPI-5482)
Campylobacter jejuni subsp. jejuni serotype O:2 (strain NCTC 11168)
Coxiella burnetii (strain RSA 493 / Nine Mile phase I)
Escherichia coli (strain K12)
Helicobacter pylori (strain ATCC 700392 / 26695)
Helicobacter pylori (strain ATCC 700392 / 26695)
Helicobacter pylori (strain ATCC 700392 / 26695)
Helicobacter pylori (strain ATCC 700392 / 26695)
Helicobacter pylori (strain ATCC 700392 / 26695)
Helicobacter pylori (strain ATCC 700392 / 26695)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
13000
-
-
gel filtration
18580
-
-
calculated from amino acid sequence
19000
-
-
isoenzyme SK1, gel filtration
20000
-
-, P56073
SDS-PAGE
21400
-
-
isoenzyme SK2, gel filtration
28000
-
-
gel filtration
additional information
-
-
aggregate containing 5 activities: EC 1.1.1.25, EC 2.7.1.71, EC 4.2.1.10, ED 4.2.3.4 and EC 2.5.1.19 has a MW of 270000 Da determined by glycerol-density-gradient centrifugation
additional information
-
-
molecular weight of enzyme aggregate containing 5 activities: EC 1.1.1.25, EC 2.7.1.71, EC 4.2.1.10, EC 4.2.3.4 and EC 2.5.1.19 is 290000 Da determined by gel filtration
additional information
-
-
molecular weight of enzyme aggregate containing 5 activities: EC 1.1.1.25, EC 2.7.1.71, EC 4.2.1.10, EC 4.2.3.4 and EC 2.5.1.19 is 270000 Da determined by gel filtration
additional information
-
-
molecular weight of enzyme aggregate containing 5 activities: EC 1.1.1.25, EC 2.7.1.71, EC 4.2.1.10, EC 4.2.3.4 and EC 2.5.1.19 is 280000 Da determined by gel filtration
additional information
-
Saccharomycopsis lipolytica
-
molecular weight of enzyme aggregate containing 5 activities: EC 1.1.1.25, EC 2.7.1.71, EC 4.2.1.10, EC 4.2.3.4 and EC 2.5.1.19 is 290000 Da determined by gel filtration
additional information
-
-
molecular weight of enzyme aggregate containing 5 activities: EC 1.1.1.25, EC 2.7.1.71, EC 4.2.1.10, EC 4.2.3.4 and EC 2.5.1.19 is 140000-145000 Da determined by gel filtration or glycerol density gradient centrifugation
additional information
-
-
MW calculated from nucleotide sequence of the pentafunctional arom polypeptide is 174555 Da
additional information
-
-
identification of the gene
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 26973.6, sequence calculation
?
Shigella flexneri 8401
-
x * 26973.6, sequence calculation
-
dimer
P56073
detailed structure-activity relationship analysis, overview
monomer
-
1 * 17000, isoenzyme SK2, SDS-PAGE
monomer
-
1 * 31000, SDS-PAGE
monomer
Escherichia coli K12
-
1 * 17000, isoenzyme SK2, SDS-PAGE
-
additional information
-
aggregate containing 5 activities: EC 1.1.1.25, EC 2.7.1.71, EC 4.2.1.10, EC 4.2.3.4 and EC 2.5.1.19 is composed of two subunits of 165000 Da
additional information
-
95% of the functional enzyme system in crude extracts exists in a dimeric form and both polypeptide chains of the homodimer are required for full activity of each of the five enzymes
additional information
-
prediction of enzyme secondary sstructure, overview
additional information
-, Q8GY88
AtSK2 adopts an alpha-beta-alpha fold with a central sheet of five parallel beta-strands flanked by two layers of alpha-helices. The AtSK2 structure can be divided into four domains: the reduced CORE domain comprising the central beta-sheet and flanking alpha-helices, the nucleotide binding domain, which includes the phosphate binding loop (P-loop/Walker-A motif), the disordered LID domain, which contains catalytic and substrate binding residues, and the extended shikimate binding domain, which includes a modified SK-type Walker B motif and several other substrate binding residues. Computational analysis of isozyme AtSK2 structure, overview
additional information
Shigella flexneri 8401
-
prediction of enzyme secondary sstructure, overview
-
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
proteolytic modification
-, Q5NTH3, Q5NTH4, Q7X7H9
protein import into intact chloroplasts isolated from Pisum sativum seedlings reveals that the full-length form is translocated into chloroplasts and processed. NH2 -terminal sequence functions as chloroplast transit peptide; protein import into intact chloroplasts isolated from Pisum sativum seedlings reveals that the full-length form is translocated into chloroplasts and processed. NH2 -terminal sequence functions as chloroplast transit peptide; protein import into intact chloroplasts isolated from Pisum sativum seedlings reveals that the full-length form is translocated into chloroplasts and processed. NH2 -terminal sequence functions as chloroplast transit peptide
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
heat-stable isozyme AtSK2, X-ray diffraction structure determinaion and analysis at 2.35 A resolution
-, Q8GY88
structural model based on structure of Mycobacterium tuberculosis enzyme. Molecular modeling and molecular dynamics simulation. Substrate shikimate binds to a pocket formed by the conserved residues Asp33, Arg57, Gly78 and Gly79, and Arg135
-
mutant enzyme K15M, sitting-drop vapor diffusion
-
vapor-diffusion method using NaCl as precipitant
-
hanging-drop vapor diffusion method. 1.8 A crystal structure of shikimate kinase. The crystal structure shows a three-layer alpha/beta fold consisting of a central sheet of five parallel beta-strands flanked by seven alpha-helices. An HpSK-shikimate-PO4 complex is also determined and refined to 2.3 A, revealing induced-fit movement from an open to a closed form on substrate binding; hanging drop vapour diffusion method using containing 0.2 M lithium sulfate, 30% (wt/vol) PEG 8000, and 0.1 M sodium acetate buffer (pH 6.5), at 20C
-, P56073
molecular modeling and docking of inhibitors. The active site is rather roomy and deep, forming an L-shape channel on the surface of the protein, and compound 3-methoxy-4-[[2-([2-methoxy-4-[(4-oxo-2-thioxo-1,3-thiazolidin-5-ylidene)methyl]phenoxy]methyl)benzyl]oxy]benzaldehyde prefers the corner area of L-shape channel, while compound 5-bromo-2-(5-[[1-(3,4-dichlorophenyl)-3,5-dioxo-4-pyrazolidinylidene]methyl]-2-furyl)benzoic acid binds the short arm of the channel in the binding interactions
-
wild-type dimeric enzyme, wild-type enzyme in complex with products ADP and shikimate 3-phosphate, enzyme mutant R57A, and enzyme mutant E114A in complex with selective inhibitor NSC162535, hanging drop vapour ddiffusion metho, 50 mg/ml protein in 40 mM Tris-HCl, pH 7.0, containing 100 mM NaCl mixed with an equal volume of reservoir solution and equilibrated against 0.06 ml of reservoir solution, containing 0.2 M Li2SO4, 30% w/v PEG 8000, and 0.1 M sodium acetate, pH 6.5 for the apo-enzyme, or containing 0.1 M HEPES sodium salt, pH 7.5, 0.1 M sodium acetate, 18% w/v PEG 8000, 2% w/v 2-propanol, and 5 mM shikimate and 5 mM MgATP for the product complex enzyme, or containing 0.1 M HEPES sodium salt, pH 8.0, 8% w/v 2-propanol and 18% w/v PEG 4000 for enzyme mutant R57A, or containing 0.1 M HEPES sodium salt, pH 6.7, and 1.2 M potassium sodium tartrate tetrahydrate for the enzyme mutant E114A with inhibitor, X-ray diffraction structure determination and analysis at 1.8 A, 2.3 A, 2.4 A, and 2.53 A resolution, respectively, molecular replacement
P56073
crystal structure of the enzyme complexed with MgADP and shikimic acid, determined at 2.3 A resolution, hanging-drop vapor-diffusion method
-
crystal structure of the enzyme in complex with MgADP- determined at 1.8 A resolution
-
hanging drop vapour diffusion method
-
hanging drop vapour diffusion method, enzyme in complex with ADP and shikimate in the absence of Mg2+ using 0.1 M Tris-HCl buffer pH 8.0, 17% PEG 1500 and 0.5-0.7 M LiCl, or enzyme in complex with ADP and Mg2+ in the absence of shikimate using 0.1 M Tris-HCl buffer pH 8.0, 20% PEG 3350 and 0.1 M MgCl2*6H2O; in complex with ADP-shikimate and with MgADP-, at 1.93 A and 2.8 A resolution, respectively. Presence of Mg2+ influences the conformation of the shikimate hydroxyl groups and the position of the side chains of some of the residues of the activesite. Presence of Cl- seems to influence the affinity of ADP and its position in the active site and the opening length of the LID domain. Shikimate binding causes a closing of the LID domain and also seems to influence the crystallographic packing
-
hanging-drop vapour-diffusion method. Crystal structure of shikimate kinase complexed with MgADP and shikimate determined at 2.3 A resolution
-
sitting drop vapour diffusion method using 0.2 M ammonium sulfate and 30% (w/v) poly(ethylene glycol) monomethyl ether 5000 in 0.1 M MES (pH 6.5) buffer
-
X-ray crystal structure of shikimate kinase with bound shikimate and adenosine diphosphate determined to a resolution of 2.15 A, sitting drop vapor diffusion method
-
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-, Q8GY88
isozyme AtSK2 is highly unstable and becomes inactivated at 37C, whereas the heat-induced isoform, AtSK1, is thermostable and fully active at 37C
40
-
-
midpoint of protein unfolding transition is 39.7C for the wild-type enzyme, 39C for the mutant enzyme C13S and 43.0C for the wild-type enzyme K15M in absence of ligands
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
the enzyme is fully unfolded in 4 M urea
-
unfolding of the enzyme by guanidinium chloride, in the absence of ligands there is a loss of structure over the range of 1-3 M guanidinium chloride
-
differential scanning calorimetry experiments for evaluaton of the stability and unfolding of each of the enzyme mutants, overview
P56073
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20C, protein concentration 1 mg/ml in 0.05% M Tris-HCl buffer, pH 7.5, 10 mM MgCl2, 0.1 M NaCl, 1 mM DTT, 10% loss of activity after 1 week, 90% loss of activity after 2 months
-
-20C, 50% glycerol, aggregate containing 5 activities: EC 1.1.1.25, EC 2.7.1.71, EC 4.2.1.10, 5-dehydroquinate synthase and 3-enoyl-pyruvylshikimate 5-phosphate synthase is stable for at least 1 month
-
4C, 50% glycerol, aggregate containing 5 activities: EC 1.1.1.25, EC 2.7.1.71, EC 4.2.1.10, 5-dehydroquinate synthase and 3-enoyl-pyruvylshikimate 5-phosphate synthase is stable for at least 19 days
-
-18C, 30% v/v glycerol, stable for several months with gradual loss of activity
-
-20C, in presence of 15% glycerol, stable for up to 4 weeks without loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
isoenzyme SK1 and SK2
-
shikimate kinase II
-
immobilized-nickel ion affinity chromatography and Sephacryl S-100 gel filtration
-
immobilized-nickel ion chromatography and Superdex-75 gel filtration
-, P56073
recombinant N-termminally His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 (DE3) by immobilized metal affinity chromatography and dialysis
-
enzyme aggregate contains 5 activities: EC 1.1.1.25, EC 2.7.1.71, EC 4.2.1.10, EC 4.2.3.4 and EC 2.5.1.19
-
partial
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
gene expression patterns of isoforms, AtSK1 and AtSK2 showing a signature of regulatory subfunctionalization
-, Q8GY88
wild-type and mutant enzyme expressed in Escherichia coli
-
cloning of aroK encoding shikimate kinase I. AroK protein, i.e. shikimate kinase I and AroL protein, i.e. shikimate kinase II are of comparable length and the homology between them extends the entire length of the two enzymes
-
generation of knockout and antisense strain from Escherichia coli parent strain DH5alpha
-
expressed in Escherichia coli BL21(DE3) cells
-
expressed in Escherichia coli JM109 cells
-, P56073
protein is expressed in Escherichia coli as N-terminal fusion with His6 tag and a TEV-protease cleavage site
-
aroK-endoded shikimate kinase, cloned and overexpressed in soluble form in Escherichia coli
-
expressed in Escherichia coli strain BL21(DE3)
-
expression of N-termminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3)
-
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression of plant shikimate kinase is induced under specific conditions of environmental stress and developmental requirements in an isoform-specific manner
-, Q8GY88
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
K138N
-, Q8GY88
AtSK1 mutation relative to AtSK2 predicted to confer substantial stabilizing effect
K87R
-, Q8GY88
AtSK1 mutation relative to AtSK2 predicted to confer substantial stabilizing effect
L233F
-, Q8GY88
AtSK1 mutation relative to AtSK2 predicted to confer substantial stabilizing effect
N234K
-, Q8GY88
AtSK1 mutation relative to AtSK2 predicted to confer substantial stabilizing effect
R120K
-, Q8GY88
AtSK1 mutation relative to AtSK2 predicted to confer substantial stabilizing effect
S152N
-, Q8GY88
AtSK1 mutation relative to AtSK2 predicted to confer substantial stabilizing effect
S251N
-, Q8GY88
AtSK1 mutation relative to AtSK2 predicted to confer substantial stabilizing effect
T230S
-, Q8GY88
AtSK1 mutation relative to AtSK2 predicted to confer substantial stabilizing effect
C13S
-
enzymatically active mutant, turnover-number is 65% of that of the wild-type enzyme
C162S
-
turnover-number is 1.14fold higher than that of the wild-type enzyme
D34N
-
inactive mutant enzyme
K15M
-
inactive mutant enzyme, increased thermostability and affinity for ATP when compared to the wild-type enzyme, the organization of the P-loop and flanking regions is heavily disturbed
D33A
P56073
site-directed mutagensis, inactive mutant
D33E
P56073
site-directed mutagensis, inactive mutant
E114A
P56073
site-directed mutagensis, the mutant shows 82% of wlld-type activity
F48A
P56073
site-directed mutagensis, inactive mutant
F48Y
P56073
site-directed mutagensis, the mutant shows 40% of wlld-type activity
M10A
P56073
site-directed mutagensis, the mutant shows 38% of wlld-type activity
R116A
P56073
site-directed mutagensis, inactive mutant
R116K
P56073
site-directed mutagensis, inactive mutant
R132A
P56073
site-directed mutagensis, the mutant shows 5% of wlld-type activity
R132K
P56073
site-directed mutagensis, inactive mutant
R57A
P56073
site-directed mutagensis, the mutant shows 2% of wlld-type activity
R57K
P56073
site-directed mutagensis, the mutant shows 2% of wlld-type activity
K15I
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
K15R
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
R110A
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
R117
-
site-directed mutagenesis, inactive mutant
T17I
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
T17R
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
K15M
-
no detectable enzyme activity
additional information
-
inactivation of the aroK gene in an shikimate-producing Escherichia coli strain DHPYA-T7. In this strain, the aroL, ptsHIcrr and ydiB genes are deleted, and the tktA, glk, aroE and aroB genes are overexpressed. Accumulation of shikimate increases 2.69fold after aroK gene deletion and 1.29fold after antisense RNA interference. The activity of shikimate kinase in the knockout strain DHPYAAS-T7 is 0.21fold of that in the antisense strain DHPYAS-T7, while the accumulation of shikimate is 1.5fold in the knockout strain DHPYAAS-T7 compared to that in the antisense strain DHPYAS-T7
additional information
Escherichia coli DHPYA-T7
-
inactivation of the aroK gene in an shikimate-producing Escherichia coli strain DHPYA-T7. In this strain, the aroL, ptsHIcrr and ydiB genes are deleted, and the tktA, glk, aroE and aroB genes are overexpressed. Accumulation of shikimate increases 2.69fold after aroK gene deletion and 1.29fold after antisense RNA interference. The activity of shikimate kinase in the knockout strain DHPYAAS-T7 is 0.21fold of that in the antisense strain DHPYAS-T7, while the accumulation of shikimate is 1.5fold in the knockout strain DHPYAAS-T7 compared to that in the antisense strain DHPYAS-T7
-
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
the enzyme is fully unfolded in 4 M urea. Approximately 95% of the enzyme activity can be recovered on dilution of the urea from 4 to 0.36 M. Refolding occurs in at least four kinetic phases, the slowest of which corresponds with the regain of shikimate binding and enzyme activity
-
when the enzyme is unfolded by incubation in 4 M urea, addition of NaCl or Na2SO4 leads to relatively slow refolding of the enzyme. The refolded enzyme can bind shikimate, though more weakly than the native enzyme. The refolded enzyme does not appear to be capable of binding nucleotides, nor does it possess detectable catalytic activity. The refolding process brought about by addition of salt in the presence of 4 M urea is not associated with any change in the fluorescence of the probe 8-anilino-1-naphthalenesulfonic acid
-
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
synthesis
-
the aroK enzyme knockout strain of Escherichia coli is useful for shikimate production
synthesis
Escherichia coli DHPYA-T7
-
the aroK enzyme knockout strain of Escherichia coli is useful for shikimate production
-
drug development
-
the enzyme is a target for antimicrobial and anti-paarasite drugs
drug development
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the enzyme is an attractive drug target as it is vital for the survival of Mycobacterium tuberculosis but absent in mammalian hosts
drug development
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the enzyme is an attractive drug target
drug development
Shigella flexneri 8401
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the enzyme is an attractive drug target
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