BRENDA - Enzyme Database
show all sequences of 4.2.99.20

Molecular basis of the general base catalysis of an alpha/beta-hydrolase catalytic triad

Sun, Y.; Yin, S.; Feng, Y.; Li, J.; Zhou, J.; Liu, C.; Zhu, G.; Guo, Z.; J. Biol. Chem. 289, 15867-15879 (2014)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
-
Escherichia coli
Crystallization (Commentary)
Crystallization (Commentary)
Organism
to 1.45 A resolution. The nucleophilicity of the catalytic serine-histidine-aspartate triad is shielded and its catalytic role is limited to being a specific general base by an open-closed conformational change
Escherichia coli
Engineering
Protein Variants
Commentary
Organism
F153A
residue involved in open-closed transition, mutation leads to large decrease in enzymatic activity
Escherichia coli
V152A
residue involved in open-closed transition, mutation leads to large decrease in enzymatic activity
Escherichia coli
V152G
residue involved in open-closed transition, mutation leads to large decrease in enzymatic activity
Escherichia coli
V152G/F153A
inactive
Escherichia coli
W147A/Y148A
inactive
Escherichia coli
Y148A
residue involved in open-closed transition, mutation leads to large decrease in enzymatic activity
Escherichia coli
Y148F
residue involved in open-closed transition, mutation leads to large decrease in enzymatic activity
Escherichia coli
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.013
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
wild-type, pH 7.0, temperature not specified in the publication
Escherichia coli
0.036
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant Y148F, pH 7.0, temperature not specified in the publication
Escherichia coli
0.05
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant Y148A, pH 7.0, temperature not specified in the publication
Escherichia coli
0.07
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant F153A, pH 7.0, temperature not specified in the publication
Escherichia coli
0.16
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant V152G, pH 7.0, temperature not specified in the publication
Escherichia coli
0.3
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant V152A, pH 7.0, temperature not specified in the publication
Escherichia coli
Organism
Organism
UniProt
Commentary
Textmining
Escherichia coli
P37355
-
-
Reaction
Reaction
Commentary
Organism
Reaction ID
5-enolpyruvoyl-6-hydroxy-2-succinylcyclohex-3-ene-1-carboxylate = (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate + pyruvate
the nucleophilicity of the catalyitc serine-histidine-aspartate triad is shielded and its catalytic role is limited to being a specific general base by an open-closed conformational change. The enzyme adopts an open conformation without a functional triad in its ligand-free form and a closed conformation with a fully functional catalytic triad in the presence of its reaction product. The open-to-closed conformational transition involves movement of half of the alpha-helical cap domain, which causes extensive structural changes in the apha/beta-domain and forces the side chainof the triad histidine to adopt an energetically disfavored gauche conformation to form the functional triad. The inactive open conformation without a triad prevails in ligand-free solution and is converted to the closed conformation with a properly formed triad by the reaction product
Escherichia coli
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
-
730042
Escherichia coli
(1R,6R)-2-succinyl-6-hydroxy-2,4-cyclohexadiene-1-carboxylate + pyruvate
-
-
-
?
Synonyms
Synonyms
Commentary
Organism
MenH
-
Escherichia coli
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.13
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant V152A, pH 7.0, temperature not specified in the publication; mutant Y148A, pH 7.0, temperature not specified in the publication
Escherichia coli
0.2
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant V152G, pH 7.0, temperature not specified in the publication
Escherichia coli
0.58
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant Y148F, pH 7.0, temperature not specified in the publication
Escherichia coli
3.33
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant F153A, pH 7.0, temperature not specified in the publication
Escherichia coli
8.5
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
wild-type, pH 7.0, temperature not specified in the publication
Escherichia coli
Cloned(Commentary) (protein specific)
Commentary
Organism
-
Escherichia coli
Crystallization (Commentary) (protein specific)
Crystallization
Organism
to 1.45 A resolution. The nucleophilicity of the catalytic serine-histidine-aspartate triad is shielded and its catalytic role is limited to being a specific general base by an open-closed conformational change
Escherichia coli
Engineering (protein specific)
Protein Variants
Commentary
Organism
F153A
residue involved in open-closed transition, mutation leads to large decrease in enzymatic activity
Escherichia coli
V152A
residue involved in open-closed transition, mutation leads to large decrease in enzymatic activity
Escherichia coli
V152G
residue involved in open-closed transition, mutation leads to large decrease in enzymatic activity
Escherichia coli
V152G/F153A
inactive
Escherichia coli
W147A/Y148A
inactive
Escherichia coli
Y148A
residue involved in open-closed transition, mutation leads to large decrease in enzymatic activity
Escherichia coli
Y148F
residue involved in open-closed transition, mutation leads to large decrease in enzymatic activity
Escherichia coli
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.013
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
wild-type, pH 7.0, temperature not specified in the publication
Escherichia coli
0.036
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant Y148F, pH 7.0, temperature not specified in the publication
Escherichia coli
0.05
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant Y148A, pH 7.0, temperature not specified in the publication
Escherichia coli
0.07
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant F153A, pH 7.0, temperature not specified in the publication
Escherichia coli
0.16
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant V152G, pH 7.0, temperature not specified in the publication
Escherichia coli
0.3
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant V152A, pH 7.0, temperature not specified in the publication
Escherichia coli
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
-
730042
Escherichia coli
(1R,6R)-2-succinyl-6-hydroxy-2,4-cyclohexadiene-1-carboxylate + pyruvate
-
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.13
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant V152A, pH 7.0, temperature not specified in the publication; mutant Y148A, pH 7.0, temperature not specified in the publication
Escherichia coli
0.2
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant V152G, pH 7.0, temperature not specified in the publication
Escherichia coli
0.58
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant Y148F, pH 7.0, temperature not specified in the publication
Escherichia coli
3.33
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant F153A, pH 7.0, temperature not specified in the publication
Escherichia coli
8.5
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
wild-type, pH 7.0, temperature not specified in the publication
Escherichia coli
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.43
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant V152A, pH 7.0, temperature not specified in the publication
Escherichia coli
1.2
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant V152G, pH 7.0, temperature not specified in the publication
Escherichia coli
2.7
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant Y148A, pH 7.0, temperature not specified in the publication
Escherichia coli
16.3
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant Y148F, pH 7.0, temperature not specified in the publication
Escherichia coli
48
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant F153A, pH 7.0, temperature not specified in the publication
Escherichia coli
533
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
wild-type, pH 7.0, temperature not specified in the publication
Escherichia coli
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.43
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant V152A, pH 7.0, temperature not specified in the publication
Escherichia coli
1.2
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant V152G, pH 7.0, temperature not specified in the publication
Escherichia coli
2.7
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant Y148A, pH 7.0, temperature not specified in the publication
Escherichia coli
16.3
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant Y148F, pH 7.0, temperature not specified in the publication
Escherichia coli
48
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant F153A, pH 7.0, temperature not specified in the publication
Escherichia coli
533
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
wild-type, pH 7.0, temperature not specified in the publication
Escherichia coli
Other publictions for EC
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
730042
Sun
Molecular basis of the general ...
Escherichia coli
J. Biol. Chem.
289
15867-15879
2014
-
-
1
1
7
-
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6
-
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1
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1
-
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1
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1
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5
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1
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1
7
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6
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-
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1
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-
5
-
-
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-
-
-
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-
6
6
730729
Johnston
Crystal structures of E. coli ...
Escherichia coli
PLoS ONE
8
e61325
2013
-
-
-
1
-
-
-
-
-
-
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2
-
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1
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-
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714632
Dawson
Exploiting the high-resolution ...
Staphylococcus aureus
BMC Struct. Biol.
11
19
2011
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1
1
-
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-
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1
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1
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1
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1
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-
702326
Jiang
Catalytic mechanism of SHCHC s ...
Escherichia coli
Biochemistry
48
6921-6931
2009
-
-
1
-
12
-
-
13
-
-
-
-
-
1
-
-
1
-
-
-
-
-
1
-
3
-
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-
13
-
-
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1
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12
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13
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1
-
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-
1
-
-
-
-
13
-
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-
-
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-
-
-
13
13
685225
Jiang
Identification and characteriz ...
Escherichia coli
Biochemistry
47
3426-3434
2008
-
-
-
-
3
-
-
10
-
1
1
1
-
4
-
-
1
-
-
-
-
-
3
1
4
1
-
-
5
1
-
-
-
-
-
-
-
-
-
-
-
3
-
-
-
-
10
-
1
1
1
-
-
-
1
-
-
-
-
3
1
1
-
-
5
1
-
-
-
-
1
1
-
5
5
685086
Jiang
Menaquinone biosynthesis in Es ...
Escherichia coli K-12
Biochemistry
46
10979-10989
2007
-
-
-
-
-
-
-
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-
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1
-
3
-
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2
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1
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