Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | the oxidized CrPGK1 retains 25% of its maximal activity and is fully activated by reducing treatments, leading to a 4fold activation | Chlamydomonas reinhardtii |
Cloned (Comment) | Organism |
---|---|
gene PGK1, recombinant expression of wild-type and mutant His-tagged enzymes in Escherichia coli strain BL21(DE3) | Chlamydomonas reinhardtii |
Protein Variants | Comment | Organism |
---|---|---|
C108S | site-directed mutagenesis, the mutant enzyme is almost completely inactive under oxidizing conditions (DTTox), but is strongly reactivated in the presence of DTTred, suggesting that the absence of Cys108 does not alter the sensitivity of the protein to the redox environment | Chlamydomonas reinhardtii |
C227S | site-directed mutagenesis, the mutant is contitutively active and is totally insensitive to oxidizing treatments, and no variation of protein activity is observed after incubation with DTTred. Absence of Cys227 allows the mutant proteins to maintain a fully active conformation comparable with the reduced wild-type enzyme | Chlamydomonas reinhardtii |
C361S | site-directed mutagenesis, the mutant is contitutively active and is totally insensitive to oxidizing treatments, and no variation of protein activity is observed after incubation with DTTred. Absence of Cys361 allows the mutant proteins to maintain a fully active conformation comparable with the reduced wild-type enzyme | Chlamydomonas reinhardtii |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | PGK1 activity is inhibited by the formation of a single regulatory disulfide bond Cys227-Cys361 with a low midpoint redox potential | Chlamydomonas reinhardtii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics, kinetic analysis or CrPGK1, overview | Chlamydomonas reinhardtii | |
0.31 | - |
ATP | pH 7.9, 25°C, recombinant oxidized wild-type enzyme | Chlamydomonas reinhardtii | |
0.35 | - |
3-phospho-D-glycerate | pH 7.9, 25°C, recombinant oxidized wild-type enzyme | Chlamydomonas reinhardtii | |
0.37 | - |
3-phospho-D-glycerate | pH 7.9, 25°C, recombinant reduced wild-type enzyme | Chlamydomonas reinhardtii | |
0.4 | - |
ATP | pH 7.9, 25°C, recombinant reduced wild-type enzyme | Chlamydomonas reinhardtii |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
chloroplast | - |
Chlamydomonas reinhardtii | 9507 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | essentially required | Chlamydomonas reinhardtii |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
45000 | - |
- |
Chlamydomonas reinhardtii |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 3-phospho-D-glycerate | Chlamydomonas reinhardtii | - |
ADP + 3-phospho-D-glyceroyl phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Chlamydomonas reinhardtii | A8JC04 | - |
- |
Oxidation Stability | Organism |
---|---|
the oxidized CrPGK1 retains 25% of its maximal activity and is fully activated by reducing treatments, leading to a 4fold activation | Chlamydomonas reinhardtii |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Chlamydomonas reinhardtii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 3-phospho-D-glycerate | - |
Chlamydomonas reinhardtii | ADP + 3-phospho-D-glyceroyl phosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 45000, oxidized enzyme form, SDS-PAGE | Chlamydomonas reinhardtii |
More | CrPGK1 is a monomeric enzyme with two distinct domains, wild-type and mutant enzyme structures by dynamic light scattering. Peptide mass fingerprinting of CrPGK1-WT and CrPGK1-C227Smutant after trypsin digestion, overview | Chlamydomonas reinhardtii |
Synonyms | Comment | Organism |
---|---|---|
PGK1 | - |
Chlamydomonas reinhardtii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Chlamydomonas reinhardtii |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
28.5 | - |
ATP | pH 7.9, 25°C, recombinant oxidized wild-type enzyme | Chlamydomonas reinhardtii | |
28.5 | - |
3-phospho-D-glycerate | pH 7.9, 25°C, recombinant oxidized wild-type enzyme | Chlamydomonas reinhardtii | |
83.4 | - |
ATP | pH 7.9, 25°C, recombinant reduced wild-type enzyme | Chlamydomonas reinhardtii | |
84.7 | - |
3-phospho-D-glycerate | pH 7.9, 25°C, recombinant reduced wild-type enzyme | Chlamydomonas reinhardtii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.9 | - |
assay at | Chlamydomonas reinhardtii |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Chlamydomonas reinhardtii |
General Information | Comment | Organism |
---|---|---|
metabolism | PGK1 is a potentially light-modulated Calvin-Benson cycle enzyme | Chlamydomonas reinhardtii |
additional information | three-dimensional enzyme structure modeling and molecular dynamics, overview | Chlamydomonas reinhardtii |
physiological function | thioredoxin-dependent redox regulation of chloroplastic phosphoglycerate kinase via oxidoreduction of the Cys227-Cys361 disulfide bond. Based on molecular mechanics calculation, the formation of the disulfide is proposed to impose structural constraints in the C-terminal domain of the enzyme that may lower its catalytic efficiency | Chlamydomonas reinhardtii |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
81.4 | - |
3-phospho-D-glycerate | pH 7.9, 25°C, recombinant oxidized wild-type enzyme | Chlamydomonas reinhardtii | |
91.9 | - |
ATP | pH 7.9, 25°C, recombinant oxidized wild-type enzyme | Chlamydomonas reinhardtii | |
208.5 | - |
ATP | pH 7.9, 25°C, recombinant reduced wild-type enzyme | Chlamydomonas reinhardtii | |
229 | - |
3-phospho-D-glycerate | pH 7.9, 25°C, recombinant reduced wild-type enzyme | Chlamydomonas reinhardtii |