Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Pinto Torres, J.E.; Yuan, M.; Goossens, J.; Versees, W.; Caljon, G.; Michels, P.A.; Walkinshaw, M.D.; Magez, S.; Sterckx, Y.G.
    Structural and kinetic characterization of Trypanosoma congolense pyruvate kinase (2020), Mol. Biochem. Parasitol., 236, 111263 .
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.7.1.40 fructose 1,6-bisphosphate F16BP, lower activation Trypanosoma congolense
2.7.1.40 fructose 1,6-bisphosphate F16BP, lower activation Trypanosoma brucei brucei
2.7.1.40 fructose 2,6-bisphosphate F26BP, a much more potent allosteric activator of trypanosomatid PYKs compared to fructose 1,6-bisphosphate, F16BP Trypanosoma congolense
2.7.1.40 fructose 2,6-bisphosphate F26BP, a much more potent allosteric activator of trypanosomatid PYKs compared to fructose 1,6-bisphosphate, F16BP Trypanosoma brucei brucei

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.40 recombinant expression of His-tagged enzyme Trypanosoma congolense
2.7.1.40 recombinant expression of His-tagged enzyme Trypanosoma brucei brucei

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.7.1.40 purified recombinant His-tagged enzyme free and in complex with fructose 1,6-bisphosphate, hanging drop vapor diffusion method, mixing of 0.002 ml of 4 mg/ml protein solution with 0.002 ml of reservoir solution containing 100 mM ammonium citrate dibasic, and 12.5% PEG 3350, and equilibration against 0.150 ml reservoir solution, 20°C, method optimization, X-ray diffraction structure deterination and analysis, molecular replacement using the structure of TbrPYK tetramer complexed with fructose 2,6-bisphosphate (PDB ID 4HYV) as a search model, modeling Trypanosoma congolense
2.7.1.40 structure analysis of TbrPYK tetramer complexed with fructose 2,6-bisphosphate (PDB ID 4HYV) and comparison with the tetrameric structure of TcoPYK (PDB IDs 6SU1 and 6SU2) Trypanosoma brucei brucei

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.1.40 citrate citrate binds TcoPYK's active site, induces an R-state transition, and is a weak inhibitor of enzyme activity, 30% inhibition at 25 mM Trypanosoma congolense

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.40 0.33
-
 phosphoenolpyruvate recombinant enzyme, pH 7.2, 25°C, in presence of fructose 1,6-bisphosphate Trypanosoma congolense
2.7.1.40 0.38
-
 phosphoenolpyruvate recombinant enzyme, pH 7.2, 25°C, in presence of fructose 2,6-bisphosphate Trypanosoma congolense
2.7.1.40 0.39
-
 phosphoenolpyruvate recombinant enzyme, pH 7.2, 25°C, in presence of fructose 1,6-bisphosphate Trypanosoma brucei brucei
2.7.1.40 0.49
-
 phosphoenolpyruvate recombinant enzyme, pH 7.2, 25°C, in presence of fructose 2,6-bisphosphate Trypanosoma brucei brucei
2.7.1.40 0.65
-
 phosphoenolpyruvate recombinant enzyme, pH 7.2, 25°C Trypanosoma congolense
2.7.1.40 1.88
-
 phosphoenolpyruvate recombinant enzyme, pH 7.2, 25°C Trypanosoma brucei brucei

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.1.40 K+ required Trypanosoma congolense
2.7.1.40 K+ required Trypanosoma brucei brucei
2.7.1.40 Mg2+ required Trypanosoma congolense
2.7.1.40 Mg2+ required Trypanosoma brucei brucei

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.1.40 ADP + phosphoenolpyruvate Trypanosoma congolense
-
 ATP + pyruvate
-
 ?
2.7.1.40 ADP + phosphoenolpyruvate Trypanosoma brucei brucei
-
 ATP + pyruvate
-
 ?
2.7.1.40 ADP + phosphoenolpyruvate Trypanosoma congolense IL3000
-
 ATP + pyruvate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.40 Trypanosoma brucei brucei P30615
-
-
2.7.1.40 Trypanosoma congolense G0UYF4
-
-
2.7.1.40 Trypanosoma congolense IL3000 G0UYF4
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.40 recombinant His-tagged enzyme Trypanosoma congolense
2.7.1.40 recombinant His-tagged enzyme Trypanosoma brucei brucei

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.40 ADP + phosphoenolpyruvate
-
 Trypanosoma congolense ATP + pyruvate
-
 ?
2.7.1.40 ADP + phosphoenolpyruvate
-
 Trypanosoma brucei brucei ATP + pyruvate
-
 ?
2.7.1.40 ADP + phosphoenolpyruvate
-
 Trypanosoma congolense IL3000 ATP + pyruvate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.7.1.40 Pyk1
-
 Trypanosoma brucei brucei
2.7.1.40 pyruvate kinase 1
-
 Trypanosoma brucei brucei
2.7.1.40 TbrPYK
-
 Trypanosoma brucei brucei
2.7.1.40 TCIL3000_10_12020
-
 Trypanosoma congolense
2.7.1.40 TcoPYK
-
 Trypanosoma congolense

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.1.40 25
-
 assay at Trypanosoma congolense
2.7.1.40 25
-
 assay at Trypanosoma brucei brucei

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.1.40 86.83
-
 phosphoenolpyruvate recombinant enzyme, pH 7.2, 25°C Trypanosoma congolense
2.7.1.40 104.33
-
 phosphoenolpyruvate recombinant enzyme, pH 7.2, 25°C Trypanosoma brucei brucei
2.7.1.40 107.33
-
 phosphoenolpyruvate recombinant enzyme, pH 7.2, 25°C, in presence of fructose 1,6-bisphosphate Trypanosoma congolense
2.7.1.40 111.5
-
 phosphoenolpyruvate recombinant enzyme, pH 7.2, 25°C, in presence of fructose 2,6-bisphosphate Trypanosoma congolense
2.7.1.40 152
-
 phosphoenolpyruvate recombinant enzyme, pH 7.2, 25°C, in presence of fructose 2,6-bisphosphate Trypanosoma brucei brucei
2.7.1.40 185
-
 phosphoenolpyruvate recombinant enzyme, pH 7.2, 25°C, in presence of fructose 1,6-bisphosphate Trypanosoma brucei brucei

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.1.40 7.2
-
 assay at Trypanosoma congolense
2.7.1.40 7.2
-
 assay at Trypanosoma brucei brucei

General Information

EC Number General Information Comment Organism
2.7.1.40 additional information analysis of structure-function relationships of pyruvate kinases (PYKs) from trypanosomatids (Trypanosoma and Leishmania), especially of TcoPYK (Uniprot ID G0UYF4) and TbrPYK (Uniprot ID P30615), overview. Substrate binding causes several structural rearrangements across the entire PYK tetramer that involve (i) AC-core rotation of 6-8° (with residues 430-434 as a pivot point), (ii) closing of the lid domain (rotation of 30-40°), (iii) stabilization of the AA' dimer interfaces, and (iv) flipping of the Arg311 side chain as part of remodeling the catalytic pocket for substrate accommodation Trypanosoma congolense
2.7.1.40 additional information analysis of structure-function relationships of pyruvate kinases (PYKs) from trypanosomatids (Trypanosoma and Leishmania), especially of TcoPYK (Uniprot ID G0UYF4) and TbrPYK (Uniprot ID P30615), overview. Substrate binding causes several structural rearrangements across the entire PYK tetramer that involve (i) AC-core rotation of 6-8° (with residues 430-434 as a pivot point), (ii) closing of the lid domain (rotation of 30-40°), (iii) stabilization of the AA' dimer interfaces, and (iv) flipping of the Arg311 side chain as part of remodeling the catalytic pocket for substrate accommodation Trypanosoma brucei brucei
2.7.1.40 physiological function allosteric regulation of trypanosomatid PYKs, overview Trypanosoma congolense
2.7.1.40 physiological function allosteric regulation of trypanosomatid PYKs, overview Trypanosoma brucei brucei

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.7.1.40 55.5
-
 phosphoenolpyruvate recombinant enzyme, pH 7.2, 25°C Trypanosoma brucei brucei
2.7.1.40 133.59
-
 phosphoenolpyruvate recombinant enzyme, pH 7.2, 25°C Trypanosoma congolense
2.7.1.40 293.42
-
 phosphoenolpyruvate recombinant enzyme, pH 7.2, 25°C, in presence of fructose 2,6-bisphosphate Trypanosoma congolense
2.7.1.40 310.2
-
 phosphoenolpyruvate recombinant enzyme, pH 7.2, 25°C, in presence of fructose 2,6-bisphosphate Trypanosoma brucei brucei
2.7.1.40 325.25
-
 phosphoenolpyruvate recombinant enzyme, pH 7.2, 25°C, in presence of fructose 1,6-bisphosphate Trypanosoma congolense
2.7.1.40 474.36
-
 phosphoenolpyruvate recombinant enzyme, pH 7.2, 25°C, in presence of fructose 1,6-bisphosphate Trypanosoma brucei brucei