BRENDA - Enzyme Database
show all sequences of 4.1.2.52

The role of a conserved histidine residue in a pyruvate-specific class II aldolase

Wang, W.; Seah, S.Y.; FEBS Lett. 582, 3385-3388 (2008)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
mutant HpaI enzymes expressed in Escherichia coli Bl21(lambda DE3) cells
Escherichia coli
Engineering
Protein Variants
Commentary
Organism
H45A
by site-directed mutagenesis, mutant enzyme has 24fold higher dissociation constant for Co2+ compared to the wild-type enzyme, apparent Km value for Co2+ increases by about 800fold compared to the wild-type enzyme, different kcat and km values
Escherichia coli
H45Q
by site-directed mutagenesis, mutant enzyme has 69fold higher dissociation constant for Co2+ compared to the wild-type enzyme, apparent Km value for Co2+ increases by about 800fold compared to the wild-type enzyme, different kcat and km values
Escherichia coli
General Stability
General Stability
Organism
the engineering results show that His45 has a structural and catalytic role. It is important for metal cofactor binding, possibly by proper positioning of a metal cofactor water ligand and is also involved in base catalysis.
Escherichia coli
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
apparent Km values for Co2+ increase in the two mutants H45A and H45Q by about 800fold compared to the wild-type enzyme
Escherichia coli
0.38
-
4-hydroxy-2-oxopentanoic acid
wild-type, pH 8.0, 25C
Escherichia coli
16
-
4-hydroxy-2-oxopentanoic acid
H45Q mutant enzyme, pH 8.0, 25C
Escherichia coli
38
-
4-hydroxy-2-oxopentanoic acid
H45A mutant enzyme, pH 8.0, 25C
Escherichia coli
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
additional information
H45A and H45Q mutant enzymes have 24fold and 69fold higher dissociation constants for Co2+ compared to the wild-type enzyme
Escherichia coli
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
additional information
-
sizes of the native mutant enzymes are identical to that of the wild-type enzyme as determined by gel filtration
Escherichia coli
Organism
Organism
UniProt
Commentary
Textmining
Escherichia coli
Q47098
-
-
Purification (Commentary)
Purification (Commentary)
Organism
by anion exchange, hydrophobic interaction, and gel filtration chormatography
Escherichia coli
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
4-hydroxy-2-oxopentanoic acid
-
722258
Escherichia coli
pyruvate + ?
-
-
-
?
Synonyms
Synonyms
Commentary
Organism
HpaI
-
Escherichia coli
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.17
-
4-hydroxy-2-oxopentanoic acid
H45Q mutant enzyme, pH 8.0, 25C
Escherichia coli
4.5
-
4-hydroxy-2-oxopentanoic acid
H45A mutant enzyme, pH 8.0, 25C
Escherichia coli
350
-
4-hydroxy-2-oxopentanoic acid
wild-type, pH 8.0, 25C
Escherichia coli
pH Range
pH Minimum
pH Maximum
Commentary
Organism
5
9
-
Escherichia coli
Cloned(Commentary) (protein specific)
Commentary
Organism
mutant HpaI enzymes expressed in Escherichia coli Bl21(lambda DE3) cells
Escherichia coli
Engineering (protein specific)
Protein Variants
Commentary
Organism
H45A
by site-directed mutagenesis, mutant enzyme has 24fold higher dissociation constant for Co2+ compared to the wild-type enzyme, apparent Km value for Co2+ increases by about 800fold compared to the wild-type enzyme, different kcat and km values
Escherichia coli
H45Q
by site-directed mutagenesis, mutant enzyme has 69fold higher dissociation constant for Co2+ compared to the wild-type enzyme, apparent Km value for Co2+ increases by about 800fold compared to the wild-type enzyme, different kcat and km values
Escherichia coli
General Stability (protein specific)
General Stability
Organism
the engineering results show that His45 has a structural and catalytic role. It is important for metal cofactor binding, possibly by proper positioning of a metal cofactor water ligand and is also involved in base catalysis.
Escherichia coli
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
apparent Km values for Co2+ increase in the two mutants H45A and H45Q by about 800fold compared to the wild-type enzyme
Escherichia coli
0.38
-
4-hydroxy-2-oxopentanoic acid
wild-type, pH 8.0, 25C
Escherichia coli
16
-
4-hydroxy-2-oxopentanoic acid
H45Q mutant enzyme, pH 8.0, 25C
Escherichia coli
38
-
4-hydroxy-2-oxopentanoic acid
H45A mutant enzyme, pH 8.0, 25C
Escherichia coli
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
additional information
H45A and H45Q mutant enzymes have 24fold and 69fold higher dissociation constants for Co2+ compared to the wild-type enzyme
Escherichia coli
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
additional information
-
sizes of the native mutant enzymes are identical to that of the wild-type enzyme as determined by gel filtration
Escherichia coli
Purification (Commentary) (protein specific)
Commentary
Organism
by anion exchange, hydrophobic interaction, and gel filtration chormatography
Escherichia coli
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
4-hydroxy-2-oxopentanoic acid
-
722258
Escherichia coli
pyruvate + ?
-
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.17
-
4-hydroxy-2-oxopentanoic acid
H45Q mutant enzyme, pH 8.0, 25C
Escherichia coli
4.5
-
4-hydroxy-2-oxopentanoic acid
H45A mutant enzyme, pH 8.0, 25C
Escherichia coli
350
-
4-hydroxy-2-oxopentanoic acid
wild-type, pH 8.0, 25C
Escherichia coli
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
5
9
-
Escherichia coli
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.011
-
4-hydroxy-2-oxopentanoic acid
H45Q mutant enzyme, pH 8.0, 25C
Escherichia coli
0.12
-
4-hydroxy-2-oxopentanoic acid
H45A mutant enzyme, pH 8.0, 25C
Escherichia coli
930
-
4-hydroxy-2-oxopentanoic acid
wild-type, pH 8.0, 25C
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
0.011
-
4-hydroxy-2-oxopentanoic acid
H45Q mutant enzyme, pH 8.0, 25C
Escherichia coli
0.12
-
4-hydroxy-2-oxopentanoic acid
H45A mutant enzyme, pH 8.0, 25C
Escherichia coli
930
-
4-hydroxy-2-oxopentanoic acid
wild-type, pH 8.0, 25C
Escherichia coli
Other publictions for EC 4.1.2.52
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)
725493
Coincon
Crystal structure of reaction ...
Escherichia coli
J. Biol. Chem.
287
36208-36221
2012
-
-
-
1
5
-
-
4
-
1
-
1
-
1
-
-
-
-
-
-
-
-
3
1
2
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
1
5
-
-
-
-
4
-
1
-
1
-
-
-
-
-
-
-
-
3
1
-
-
-
4
-
-
-
-
-
-
-
-
3
3
702377
Wang
Comparison of two metal-depend ...
Escherichia coli
Biochemistry
49
3774-3782
2010
-
-
-
1
-
-
5
10
-
1
-
1
-
1
-
-
-
1
-
-
-
-
9
-
1
-
-
-
9
-
-
-
-
6
-
-
-
-
-
-
1
-
-
-
5
6
10
-
1
-
1
-
-
-
-
-
-
-
-
9
-
-
-
-
9
-
-
-
-
-
-
-
-
9
9
722258
Wang
The role of a conserved histid ...
Escherichia coli
FEBS Lett.
582
3385-3388
2008
-
-
1
-
2
1
-
4
-
1
1
-
-
1
-
-
1
-
-
-
-
-
1
-
1
-
-
-
3
-
1
-
-
-
-
-
-
-
1
-
-
2
1
-
-
-
4
-
1
1
-
-
-
-
1
-
-
-
-
1
-
-
-
-
3
-
1
-
-
-
-
-
-
3
3
722955
Rea
Structure and mechanism of Hpc ...
Escherichia coli
J. Mol. Biol.
373
866-876
2007
-
-
1
1
-
-
-
-
-
-
1
1
-
2
-
-
1
-
-
-
-
-
1
1
2
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
1
1
-
-
-
1
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
1
1
-
-
-
721601
Wang
Purification and biochemical c ...
Escherichia coli
Biochemistry
44
9447-9455
2005
-
-
1
-
1
-
2
6
-
9
1
1
-
1
-
-
1
-
-
-
-
-
5
-
1
-
-
-
3
-
-
-
-
2
-
-
-
-
1
-
-
1
-
-
2
2
6
-
9
1
1
-
-
-
1
-
-
-
-
5
-
-
-
-
3
-
-
-
-
-
1
1
-
3
3