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Literature summary extracted from

  • Adina-Zada, A.; Jitrapakdee, S.; Wallace, J.C.; Attwood, P.V.
    Coordinating Role of His216 in MgATP Binding and Cleavage in Pyruvate Carboxylase (2014), Biochemistry, 53, 1051-1058.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
6.4.1.1 acetyl-CoA
-
Rhizobium etli

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.4.1.1 expressed in Escherichia coli as a His-tagged fusion protein Rhizobium etli

Protein Variants

EC Number Protein Variants Comment Organism
6.4.1.1 H216N no differences to quarterny structures compared to wild-type. Mutation results in a 9fold increase in the Km for MgATP in its steady-state cleavage in the absence of pyruvate and a 3fold increase in the Km for MgADP in its steady-state phosphorylation by carbamoyl phosphate. kcat/Km (MgATP)98% decreased compared to wild-type. For MgADP phosphorylation kcat/Km is 99.5% decreased compared to wild-type. The Kd of the enzyme MgATP complex is essentially the same in the wildtype enzyme and H216N but the first-order rate constant for MgATP cleavage in the single-turnover experiments in H216N is only 0.75% of that for the wild-type enzyme, and thus, the MgATP cleavage step is rate-limiting in the steady state for H216N but not for the wild-type enzyme Rhizobium etli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.4.1.1 0.0028
-
ADP wild-type, MgADP phosphorylation, pH 7.8, 30°C Rhizobium etli
6.4.1.1 0.008
-
ADP mutant H216N, MgADP phosphorylation, pH 7.8, 30°C Rhizobium etli
6.4.1.1 0.009
-
ATP wild-type, MgATP cleavage, pH 7.8, 30°C Rhizobium etli
6.4.1.1 0.082
-
ATP mutant H216N, MgATP cleavage, pH 7.8, 30°C Rhizobium etli

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.4.1.1 Mg2+
-
Rhizobium etli

Organism

EC Number Organism UniProt Comment Textmining
6.4.1.1 Rhizobium etli Q2K340
-
-
6.4.1.1 Rhizobium etli CFN 42 Q2K340
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.4.1.1 using Ni-NTA chromatography Rhizobium etli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.4.1.1 ATP + pyruvate + HCO3-
-
Rhizobium etli ADP + phosphate + oxaloacetate
-
r
6.4.1.1 ATP + pyruvate + HCO3-
-
Rhizobium etli CFN 42 ADP + phosphate + oxaloacetate
-
r

Subunits

EC Number Subunits Comment Organism
6.4.1.1 tetramer
-
Rhizobium etli

Synonyms

EC Number Synonyms Comment Organism
6.4.1.1 RePC
-
Rhizobium etli

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.4.1.1 30
-
assay at Rhizobium etli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.4.1.1 0.052
-
pyruvate co-substrate: HCO3-, mutant H216N, pyruvate carboxylation, pH 7.8, 30°C Rhizobium etli
6.4.1.1 0.1
-
oxaloacetate co-substrate: ADP + phosphate, mutant H216N, oxaloacetate decarboxylation, pH 7.8, 30°C Rhizobium etli
6.4.1.1 0.28
-
oxaloacetate co-substrate: ADP + phosphate, wild-type, oxaloacetate decarboxylation, pH 7.8, 30°C Rhizobium etli
6.4.1.1 11
-
pyruvate co-substrate: HCO3-, wild-type, pyruvate carboxylation, pH 7.8, 30°C Rhizobium etli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.4.1.1 7.8
-
assay at Rhizobium etli

Cofactor

EC Number Cofactor Comment Organism Structure
6.4.1.1 biotin
-
Rhizobium etli

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
6.4.1.1 0.14
-
ATP mutant H216N, MgATP cleavage, pH 7.8, 30°C Rhizobium etli
6.4.1.1 2.7
-
ADP mutant H216N, MgADP phosphorylation, pH 7.8, 30°C Rhizobium etli
6.4.1.1 6.86
-
ATP wild-type, MgATP cleavage, pH 7.8, 30°C Rhizobium etli
6.4.1.1 589
-
ADP wild-type, MgADP phosphorylation, pH 7.8, 30°C Rhizobium etli