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Literature summary for 2.7.2.10 extracted from

  • Encalada, R.; Rojo-Dominguez, A.; Rodriguez-Zavala, J.S.; Pardo, J.P.; Quezada, H.; Moreno-Sanchez, R.; Saavedra, E.
    Molecular basis of the unusual catalytic preference for GDP/GTP in Entamoeba histolytica 3-phosphoglycerate kinase (2009), FEBS J., 276, 2037-2047.
    View publication on PubMed
Search for more literature for 2.7.2.10

Cloned(Commentary)

Cloned (Comment) Organism
mutant proteins cloned into the pGEM-T-easy vector, further cloned into the pET28 expression vector, wild-type and mutants overexpressed as N-terminal histidine-tailed recombinant proteins in Escherichia coli BL21DE3pLysS cells Entamoeba histolytica

Protein Variants

Protein Variants Comment Organism
E309M increases catalytic efficiency (Vm/Km) with ADP/ATP as a result of both, increased Vm and decreased Km values Entamoeba histolytica
E309Q increases catalytic efficiency (Vm/Km) with ADP/ATP as a result of both, increased Vm and decreased Km values Entamoeba histolytica
V311L increases catalytic efficiency (Vm/Km) with ADP/ATP as a result of both, increased Vm and decreased Km values. Is monomeric as the wild-type Entamoeba histolytica
Y239F increases catalytic efficiency (Vm/Km) with ADP/ATP as a result of both, increased Vm and decreased Km values Entamoeba histolytica
Y239F/E309M higher catalytic efficiency in the double mutant is achieved mainly due to an increased affinity for ADP/ATP with a concomitant diminished affinity for GDP/GTP. Is monomeric as the wild-type Entamoeba histolytica

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.016
-
 GTP wild-type, in cytosolic-enriched cellular extracts, at 37°C, pH 7.0 Entamoeba histolytica
0.07
-
 GDP wild-type, in cytosolic-enriched cellular extracts, at 37°C, pH 7.0 Entamoeba histolytica
0.108
-
 GTP recombinant purified wild-type, at 37°C, pH 7.0 Entamoeba histolytica
0.112
-
 GDP recombinant purified wild-type, at 37°C, pH 7.0 Entamoeba histolytica
0.115
-
 GTP recombinant purified mutant V311L, at 37°C, pH 7.0 Entamoeba histolytica
0.155
-
 GDP recombinant purified mutant Y239F, at 37°C, pH 7.0 Entamoeba histolytica
0.158
-
 GDP recombinant purified mutant E309Q, at 37°C, pH 7.0 Entamoeba histolytica
0.167
-
 GTP recombinant purified mutant Y239F, at 37°C, pH 7.0 Entamoeba histolytica
0.168
-
 GDP recombinant purified mutant V311L, at 37°C, pH 7.0 Entamoeba histolytica
0.169
-
 GTP recombinant purified mutant E309M, at 37°C, pH 7.0 Entamoeba histolytica
0.208
-
 GDP recombinant purified mutant E309M, at 37°C, pH 7.0 Entamoeba histolytica
0.3
-
 GTP recombinant purified mutant E309Q, at 37°C, pH 7.0 Entamoeba histolytica
0.348
-
 GTP recombinant purified mutant Y239F/E309M, at 37°C, pH 7.0 Entamoeba histolytica
0.802
-
 GDP recombinant purified mutant Y239F/E309M, at 37°C, pH 7.0 Entamoeba histolytica

Localization

Localization Comment Organism GeneOntology No. Textmining
cytosol
-
 Entamoeba histolytica 5829
-

Metals/Ions

Metals/Ions Comment Organism Structure
MgCl2
-
 Entamoeba histolytica

Organism

Organism UniProt Comment Textmining
Entamoeba histolytica
-
 HM1:IMSS
-
Entamoeba histolytica HM1:IMSS
-
 HM1:IMSS
-

Purification (Commentary)

Purification (Comment) Organism
wild-type and mutants purified by metal-affinity chromatography, mutants purified more than 98% Entamoeba histolytica

Storage Stability

Storage Stability Organism
-22°C, 50% glycerol (v/v), 4-7 months Entamoeba histolytica

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
GDP + 3-phospho-D-glyceroyl 1-phosphate
-
 Entamoeba histolytica GTP + 3-phospho-D-glycerate
-
 r
GDP + 3-phospho-D-glyceroyl 1-phosphate
-
 Entamoeba histolytica HM1:IMSS GTP + 3-phospho-D-glycerate
-
 r
GTP + 3-phospho-D-glycerate
-
 Entamoeba histolytica GDP + 3-phospho-D-glyceroyl 1-phosphate
-
 r
GTP + 3-phospho-D-glycerate
-
 Entamoeba histolytica HM1:IMSS GDP + 3-phospho-D-glyceroyl 1-phosphate
-
 r

Subunits

Subunits Comment Organism
monomer
-
 Entamoeba histolytica

Synonyms

Synonyms Comment Organism
3-phosphoglycerate kinase
-
 Entamoeba histolytica
PGK
-
 Entamoeba histolytica

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50
-
 wild-type, mutants V311L and Y239F/E309Q incubated for 1-12 min do not show drastic reductions in activity at 50°C. 30% of their initial activities decay after 1 min incubation at 60°C Entamoeba histolytica

Cofactor

Cofactor Comment Organism Structure
GDP displays higher activity with GDP/GTP and lower affinities for the adenine nucleotides Entamoeba histolytica
GTP displays higher activity with GDP/GTP and lower affinities for the adenine nucleotides Entamoeba histolytica