BRENDA - Enzyme Database

Structural role of the active-site metal in the conformation of Trypanosoma brucei phosphoglycerate mutase

Mercaldi, G.F.; Pereira, H.M; Cordeiro, A.T.; Michels, P.A.M.; Thiemann, O.H.; FEBS J. 279, 2012-2021 (2012)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
5.4.2.12
additional information
independent on 2,3-bisphosphoglycerate
Trypanosoma brucei
Application
EC Number
Application
Commentary
Organism
5.4.2.12
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(Commentary)
EC Number
Commentary
Organism
5.4.2.12
recombinant expression of the His-tagged enzyme in Escherichia coli strain BL21(DE3)
Trypanosoma brucei
Crystallization (Commentary)
EC Number
Crystallization
Organism
5.4.2.12
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
Engineering
EC Number
Amino acid exchange
Commentary
Organism
5.4.2.12
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]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
5.4.2.12
0.148
-
3-phospho-D-glycerate
recombinant enzyme, pH 7.4, 37°C
Trypanosoma brucei
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
5.4.2.12
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]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
5.4.2.12
56000
-
recombinant enzyme, gel filtration
Trypanosoma brucei
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
5.4.2.12
2-phospho-D-glycerate
Trypanosoma brucei
-
3-phospho-D-glycerate
-
-
r
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
5.4.2.12
Trypanosoma brucei
Q9NG18
-
-
Purification (Commentary)
EC Number
Commentary
Organism
5.4.2.12
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography and gel filtration
Trypanosoma brucei
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
5.4.2.12
2-phospho-D-glycerate
-
727472
Trypanosoma brucei
3-phospho-D-glycerate
-
-
-
r
5.4.2.12
3-phospho-D-glycerate
-
727472
Trypanosoma brucei
2-phospho-D-glycerate
-
-
-
r
Subunits
EC Number
Subunits
Commentary
Organism
5.4.2.12
monomer
1 * 56000, SDS-PAGE, recombinant enzyme
Trypanosoma brucei
5.4.2.12
More
ligand-induced conformational changes, overview
Trypanosoma brucei
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
5.4.2.12
37
-
assay at
Trypanosoma brucei
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
5.4.2.12
19.6
-
3-phospho-D-glycerate
recombinant enzyme, pH 7.4, 37°C
Trypanosoma brucei
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
5.4.2.12
7.4
-
assay at
Trypanosoma brucei
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
5.4.2.12
additional information
independent on 2,3-bisphosphoglycerate
Trypanosoma brucei
Application (protein specific)
EC Number
Application
Commentary
Organism
5.4.2.12
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(Commentary) (protein specific)
EC Number
Commentary
Organism
5.4.2.12
recombinant expression of the His-tagged enzyme in Escherichia coli strain BL21(DE3)
Trypanosoma brucei
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
5.4.2.12
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
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
5.4.2.12
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] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
5.4.2.12
0.148
-
3-phospho-D-glycerate
recombinant enzyme, pH 7.4, 37°C
Trypanosoma brucei
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
5.4.2.12
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] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
5.4.2.12
56000
-
recombinant enzyme, gel filtration
Trypanosoma brucei
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
5.4.2.12
2-phospho-D-glycerate
Trypanosoma brucei
-
3-phospho-D-glycerate
-
-
r
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
5.4.2.12
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography and gel filtration
Trypanosoma brucei
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
5.4.2.12
2-phospho-D-glycerate
-
727472
Trypanosoma brucei
3-phospho-D-glycerate
-
-
-
r
5.4.2.12
3-phospho-D-glycerate
-
727472
Trypanosoma brucei
2-phospho-D-glycerate
-
-
-
r
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
5.4.2.12
monomer
1 * 56000, SDS-PAGE, recombinant enzyme
Trypanosoma brucei
5.4.2.12
More
ligand-induced conformational changes, overview
Trypanosoma brucei
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
5.4.2.12
37
-
assay at
Trypanosoma brucei
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
5.4.2.12
19.6
-
3-phospho-D-glycerate
recombinant enzyme, pH 7.4, 37°C
Trypanosoma brucei
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
5.4.2.12
7.4
-
assay at
Trypanosoma brucei
General Information
EC Number
General Information
Commentary
Organism
5.4.2.12
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
5.4.2.12
additional information
ligand-induced conformational changes, overview
Trypanosoma brucei
General Information (protein specific)
EC Number
General Information
Commentary
Organism
5.4.2.12
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
5.4.2.12
additional information
ligand-induced conformational changes, overview
Trypanosoma brucei
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
5.4.2.12
3226
-
3-phospho-D-glycerate
recombinant enzyme, pH 7.4, 37°C
Trypanosoma brucei
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
5.4.2.12
3226
-
3-phospho-D-glycerate
recombinant enzyme, pH 7.4, 37°C
Trypanosoma brucei