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Literature summary for 6.4.1.1 extracted from

  • Duangpan, S.; Jitrapakdee, S.; Adina-Zada, A.; Byrne, L.; Zeczycki, T.N.; St Maurice, M.; Cleland, W.W.; Wallace, J.C.; Attwood, P.V.
    Probing the catalytic roles of Arg548 and Gln552 in the carboxyl transferase domain of the Rhizobium etli pyruvate carboxylase by site-directed mutagenesis (2010), Biochemistry, 49, 3296-3304.
    View publication on PubMedView publication on EuropePMC
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Activating Compound

Activating Compound Comment Organism Structure
acetyl-CoA
-
 Rhizobium etli

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Rhizobium etli

Protein Variants

Protein Variants Comment Organism
Q552A the mutation results in loss of the ability to catalyse pyruvate carboxylation, biotin-dependent decarboxylation of oxaloacetate and proton exchange between pyruvate and water Rhizobium etli
Q552N the mutation results in loss of the ability to catalyse pyruvate carboxylation, biotin-dependent decarboxylation of oxaloacetate and proton exchange between pyruvate and water Rhizobium etli
R548A the mutation results in loss of the ability to catalyse pyruvate carboxylation, biotin-dependent decarboxylation of oxaloacetate and proton exchange between pyruvate and water Rhizobium etli
R548K the mutation results in loss of the ability to catalyse pyruvate carboxylation (2% residual activity), biotin-dependent decarboxylation of oxaloacetate and proton exchange between pyruvate and water Rhizobium etli

Inhibitors

Inhibitors Comment Organism Structure
oxamate
-
 Rhizobium etli

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required for activity Rhizobium etli

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
472000
-
 analytical ultracentrifugation Rhizobium etli

Organism

Organism UniProt Comment Textmining
Rhizobium etli
-
-
-

Purification (Commentary)

Purification (Comment) Organism
HisPur cobalt resin column chromatography Rhizobium etli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + pyruvate + HCO3- + H+
-
 Rhizobium etli ADP + oxaloacetate + phosphate
-
 r

Subunits

Subunits Comment Organism
homotetramer analytical ultracentrifugation Rhizobium etli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.28
-
 pyruvate mutant enzyme R548K, in 100 mM Tris-HCl pH 7.8, 30°C Rhizobium etli
13.9
-
 pyruvate wild type enzyme, in 100 mM Tris-HCl pH 7.8, 30°C Rhizobium etli

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Rhizobium etli
biotin
-
 Rhizobium etli

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
8
-
 oxamate mutant enzyme Q552N, in 100 mM Tris-HCl pH 7.8, 30°C Rhizobium etli
10
-
 oxamate mutant enzyme R548K, in 100 mM Tris-HCl pH 7.8, 30°C Rhizobium etli
19
-
 oxamate wild type enzyme, in 100 mM Tris-HCl pH 7.8, 30°C Rhizobium etli