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Literature summary for 2.7.2.3 extracted from

  • McHarg, J.; Kelly, S.M.; Price, N.C.; Cooper, A.; Littlechild, J.A.
    Site-directed mutagenesis of proline 204 in the 'hinge' region of yeast phosphoglycerate kinase (1999), Eur. J. Biochem., 259, 939-945.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
overexpression in Saccharomyces cerevisiae Saccharomyces cerevisiae

Protein Variants

Protein Variants Comment Organism
P204F site-directed mutagenesis, mutation in the hinge region of the enzyme, which plays a role in protein folding during catalysis, less well folded with considerable loss of secondary and tertiary structure, no activity Saccharomyces cerevisiae
P204H site-directed mutagenesis, mutation in the hinge region of the enzyme, which plays a role in protein folding during catalysis, secondary and tertiary structure is similar to the wild-type, but the mutant is less stable to heat and guanidinium chloride denaturation, 3-4fold increase of Km for 3-phospho-D-glycerate and ATP Saccharomyces cerevisiae

General Stability

General Stability Organism
Pro204 is important for stability and catalytic mechanism of the enzyme Saccharomyces cerevisiae

Inhibitors

Inhibitors Comment Organism Structure
Guanidinium chloride 0.5 M, 30% loss of activity for the mutant P204H, 5% loss of activity for the wild-type, both are unfolded at 1 M Saccharomyces cerevisiae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetics Saccharomyces cerevisiae
0.33
-
ATP wild-type enzyme, pH 7.5, 25°C Saccharomyces cerevisiae
0.77
-
3-phospho-D-glycerate wild-type enzyme, pH 7.5, 25°C Saccharomyces cerevisiae
1.25
-
ATP mutant P204H, pH 7.5, 25°C Saccharomyces cerevisiae
2.5
-
3-phospho-D-glycerate mutant P204H, pH 7.5, 25°C Saccharomyces cerevisiae

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ADP + 3-phospho-D-glyceroyl phosphate Saccharomyces cerevisiae
-
ATP + 3-phospho-D-glycerate
-
r

Organism

Organism UniProt Comment Textmining
Saccharomyces cerevisiae
-
-
-

Reaction

Reaction Comment Organism Reaction ID
ATP + 3-phospho-D-glycerate = ADP + 3-phospho-D-glyceroyl phosphate Pro204 is important for stability and catalytic mechanism of the enzyme Saccharomyces cerevisiae

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.4
-
purified mutant P204F Saccharomyces cerevisiae
4.5
-
purified mutant P204H Saccharomyces cerevisiae
468
-
purified wild-type enzyme Saccharomyces cerevisiae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ADP + 3-phospho-D-glyceroyl phosphate
-
Saccharomyces cerevisiae ATP + 3-phospho-D-glycerate
-
r
ATP + 3-phospho-D-glycerate
-
Saccharomyces cerevisiae ADP + 1,3-diphosphoglycerate
-
r

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Saccharomyces cerevisiae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.4
-
ATP mutant P204H, pH 7.5, 25°C Saccharomyces cerevisiae
3.4
-
3-phospho-D-glycerate mutant P204H, pH 7.5, 25°C Saccharomyces cerevisiae
354
-
ATP wild-type enzyme, pH 7.5, 25°C Saccharomyces cerevisiae
354
-
3-phospho-D-glycerate wild-type enzyme, pH 7.5, 25°C Saccharomyces cerevisiae

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Saccharomyces cerevisiae

Cofactor

Cofactor Comment Organism Structure
ADP
-
Saccharomyces cerevisiae
ATP required as phosphate donor Saccharomyces cerevisiae