BRENDA - Enzyme Database

Effects of the G376E and G157D mutations on the stability of yeast enolase - a model for human muscle enolase deficiency

Zhao, S.; Choy, B.S.; Kornblatt, M.J.; FEBS J. 275, 97-106 (2007)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
4.2.1.11
inserted into pET-3a, expressed in Escherichia coli BL21-DE3, site-directed mutagenesis performed
Saccharomyces cerevisiae
Engineering
EC Number
Amino acid exchange
Commentary
Organism
4.2.1.11
G157D
correctly folded, less stable than wild-type enolase, dissociation into subunit forms accelerated
Saccharomyces cerevisiae
4.2.1.11
G376E
correctly folded, less stable than wild-type enolase, dissociation into subunit forms accelerated
Saccharomyces cerevisiae
4.2.1.11
additional information
folding studies, dissociation experiments, determination of thermal and enzymatic stability
Saccharomyces cerevisiae
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
4.2.1.11
2-phospho-D-glycerate
Saccharomyces cerevisiae
-
phosphoenolpyruvate + H2O
-
-
r
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
4.2.1.11
Saccharomyces cerevisiae
P00924
-
-
Purification (Commentary)
EC Number
Commentary
Organism
4.2.1.11
recombinant proteins, SDS-PAGE
Saccharomyces cerevisiae
Source Tissue
EC Number
Source Tissue
Commentary
Organism
Textmining
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
4.2.1.11
additional information
-
enzyme activity monitored by following the conversion of phosphoenolpyruvate to 2-phospho-D-glycerate, specific activities of the variants, relative to wildtype enolase, are 0.1% for G157D and 0.01% for G376E
Saccharomyces cerevisiae
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4.2.1.11
2-phospho-D-glycerate
-
679807
Saccharomyces cerevisiae
phosphoenolpyruvate + H2O
-
-
-
r
4.2.1.11
2-phospho-D-glycerate
influence on exchange of amino acid residues on structure, dissociation and function of enolase analyzed
679807
Saccharomyces cerevisiae
phosphoenolpyruvate + H2O
-
-
-
r
Subunits
EC Number
Subunits
Commentary
Organism
4.2.1.11
More
subunit dissociation of wild-type and G157D enolases by incubation with NaClO4 at 15°C for 24 h, spectrometrically measured
Saccharomyces cerevisiae
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
4.2.1.11
additional information
-
thermal stability lower in mutant forms of enolase, wild-type enolase has Tm of 55.4°C, whereas G367E and G157D reveals a Tm of 51.2°C and 49.9°C, respectively, addition of 50 microM phosphonoacetohydroxamate increases thermal stabilization of wildtype and G157D enolases, but not of G367E
Saccharomyces cerevisiae
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
4.2.1.11
7.1
-
activity assay at
Saccharomyces cerevisiae
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
4.2.1.11
inserted into pET-3a, expressed in Escherichia coli BL21-DE3, site-directed mutagenesis performed
Saccharomyces cerevisiae
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
4.2.1.11
G157D
correctly folded, less stable than wild-type enolase, dissociation into subunit forms accelerated
Saccharomyces cerevisiae
4.2.1.11
G376E
correctly folded, less stable than wild-type enolase, dissociation into subunit forms accelerated
Saccharomyces cerevisiae
4.2.1.11
additional information
folding studies, dissociation experiments, determination of thermal and enzymatic stability
Saccharomyces cerevisiae
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
4.2.1.11
2-phospho-D-glycerate
Saccharomyces cerevisiae
-
phosphoenolpyruvate + H2O
-
-
r
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
4.2.1.11
recombinant proteins, SDS-PAGE
Saccharomyces cerevisiae
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
Organism
Textmining
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
4.2.1.11
additional information
-
enzyme activity monitored by following the conversion of phosphoenolpyruvate to 2-phospho-D-glycerate, specific activities of the variants, relative to wildtype enolase, are 0.1% for G157D and 0.01% for G376E
Saccharomyces cerevisiae
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4.2.1.11
2-phospho-D-glycerate
-
679807
Saccharomyces cerevisiae
phosphoenolpyruvate + H2O
-
-
-
r
4.2.1.11
2-phospho-D-glycerate
influence on exchange of amino acid residues on structure, dissociation and function of enolase analyzed
679807
Saccharomyces cerevisiae
phosphoenolpyruvate + H2O
-
-
-
r
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
4.2.1.11
More
subunit dissociation of wild-type and G157D enolases by incubation with NaClO4 at 15°C for 24 h, spectrometrically measured
Saccharomyces cerevisiae
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
4.2.1.11
additional information
-
thermal stability lower in mutant forms of enolase, wild-type enolase has Tm of 55.4°C, whereas G367E and G157D reveals a Tm of 51.2°C and 49.9°C, respectively, addition of 50 microM phosphonoacetohydroxamate increases thermal stabilization of wildtype and G157D enolases, but not of G367E
Saccharomyces cerevisiae
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
4.2.1.11
7.1
-
activity assay at
Saccharomyces cerevisiae