Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | independent on 2,3-bisphosphoglycerate | Trypanosoma brucei |
Application | Comment | Organism |
---|---|---|
drug development | TbiPGAM is an attractive molecular target for drug development, the apoenzyme conformation described here provides opportunities for its use in structure-based drug design approaches | Trypanosoma brucei |
Cloned (Comment) | Organism |
---|---|
recombinant expression of the His-tagged enzyme in Escherichia coli strain BL21(DE3) | Trypanosoma brucei |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His-tagged enzyme, hanging drop vapour diffusion method, mixing of 0.003 ml of 8 mg/ml protein solution containing 20 mM Tris-acetate-EDTA, pH 7.4, 50 mM NaCl, 001 mM CoCl2, with 0.003 ml of reservoir solution containing 0.05 M ammonium sulfate, 0.1 M Bis-Tris, pH 6.1, and 25% w/v PEG 3350, 18°C, X-ray diffraction structure determination and analysis at 2.3 A resolution | Trypanosoma brucei |
Protein Variants | Comment | Organism |
---|---|---|
D319A | site-directed mutagenesis, substitution of the metal-binding residue Asp319 by Ala results in complete loss of independent PGAM activity | Trypanosoma brucei |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.148 | - |
3-phospho-D-glycerate | recombinant enzyme, pH 7.4, 37°C | Trypanosoma brucei |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | two metal sites (M1 and M2) containing cobalt ions are present in the phosphatase domain of the enzyme structure, interaction between Asp319 and the metal bound to the active site, contribution to the domain movement | Trypanosoma brucei |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
56000 | - |
recombinant enzyme, gel filtration | Trypanosoma brucei |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-phospho-D-glycerate | Trypanosoma brucei | - |
3-phospho-D-glycerate | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Trypanosoma brucei | Q9NG18 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography and gel filtration | Trypanosoma brucei |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-phospho-D-glycerate | - |
Trypanosoma brucei | 3-phospho-D-glycerate | - |
r | |
3-phospho-D-glycerate | - |
Trypanosoma brucei | 2-phospho-D-glycerate | - |
r |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 56000, SDS-PAGE, recombinant enzyme | Trypanosoma brucei |
More | ligand-induced conformational changes, overview | Trypanosoma brucei |
Synonyms | Comment | Organism |
---|---|---|
independent PGAM | - |
Trypanosoma brucei |
iPGAM | - |
Trypanosoma brucei |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Trypanosoma brucei |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3226 | - |
3-phospho-D-glycerate | recombinant enzyme, pH 7.4, 37°C | Trypanosoma brucei |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Trypanosoma brucei |
General Information | Comment | Organism |
---|---|---|
evolution | PGAMs that are dependent on (EC 5.4.2.11) or independent of the 2,3-bisphosphoglycerate cofactor are members of two distinct protein families | Trypanosoma brucei |
additional information | ligand-induced conformational changes, overview | Trypanosoma brucei |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3226 | - |
3-phospho-D-glycerate | recombinant enzyme, pH 7.4, 37°C | Trypanosoma brucei |