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Literature summary for 6.4.1.1 extracted from

  • Adina-Zada, A.; Sereeruk, C.; Jitrapakdee, S.; Zeczycki, T.N.; St Maurice, M.; Cleland, W.W.; Wallace, J.C.; Attwood, P.V.
    Roles of Arg427 and Arg472 in the binding and allosteric effects of acetyl CoA in pyruvate carboxylase (2012), Biochemistry, 51, 8208-8217.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 6.4.1.1

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Rhizobium etli

Protein Variants

Protein Variants Comment Organism
R427K more than 70% of mutant exist in the tetrameric form, Ka (activation value) for acetyl-CoA is 76fold higher compared to wild-type, kcat 32% of wild-type enzyme, Km (MgATP) 2.6fold higher compared to wild-type, kcat/Km value 12% of wild-type. Ka (activation value) for Mg2+ is 8fold higher compared to wild-type. Bicarbonate-dependent ATP cleavage activity: 0.4% of wild-type in the presence of acetyl-CoA/0.8% of wild-type in the absence of acetyl-CoA. ADP phosphorylation by carbamoyl phosphate: 21% of wild-type in the presence of acetyl-CoA. In the absence of acetyl-CoA activity slightly increased compared to wild-type. Mutant exhibits 6% biotin carboxylation rate of wild-type. residual activity at saturating concentrations of L-aspartate is 2fold greater than wild-type Rhizobium etli
R427S more than 70% of mutant exist in the tetrameric form, Ka (activation value) for acetyl-CoA is 15fold higher compared to wild-type, kcat 41% of wild-type enzyme, Km (MgATP) 1.5fold higher compared to wild-type, kcat/Km value 26% of wild-type. Ka (activation value) for Mg2+ is 2fold higher compared to wild-type. Bicarbonate-dependent ATP cleavage activity: 1.1% of wild-type in the presence of acetyl-CoA/5.8% of wild-type in the absence of acetyl-CoA. ADP phosphorylation by carbamoyl phosphate: 17% of wild-type in the presence of acetyl-CoA. In the absence of acetyl-CoA activity similar to wild-type. Mutant exhibits 6% biotin carboxylation rate of wild-type Rhizobium etli
R472K 45% of mutant exist in the tetrameric form, Ka (activation value) for acetyl-CoA is 252fold higher compared to wild-type, kcat 32% of wild-type enzyme, Km (MgATP) 6.7fold higher compared to wild-type, kcat/Km value 3% of wild-type. Ka (activation value) for Mg2+ is 37fold higher compared to wild-type. Bicarbonate-dependent ATP cleavage activity: 0.6% of wild-tpye in the presence of acetyl-CoA/0.9% of wild-type in the absence of acetyl-CoA. ADP phosphorylation by carbamoyl phosphate: 8% of wild-type in the presence of acetyl-CoA. In the absence of acetyl-CoA activity slightly increased compared to wild-type. Mutant exhibits 3% biotin carboxylation rate of wild-type Rhizobium etli
R472S more than 70% of mutant exist in the tetrameric form, Ka (activation value) for acetyl-CoA is 203fold higher compared to wild-type, kcat 25% of wild-type enzyme, Km (MgATP) 8.7fold higher compared to wild-type, kcat/Km value 2% of wild-type. Ka (activation value) for Mg2+ is 30fold higher compared to wild-type. Bicarbonate-dependent ATP cleavage activity: 0.6% of wild-type in the presence of acetyl-CoA/1.2% of wild-type in the absence of acetyl-CoA. ADP phosphorylation by carbamoyl phosphate: 5% of wild-type in the presence of acetyl-CoA. In the absence of acetyl-CoA activity slightly increased compared to wild-type. Mutant exhibits 2% biotin carboxylation rate of wild-type Rhizobium etli

Inhibitors

Inhibitors Comment Organism Structure
L-aspartate
-
 Rhizobium etli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.1 1 ATP wild-type, pH 7.8, 30°C Rhizobium etli
0.16
-
 ATP mutant R427S, pH 7.8, 30°C Rhizobium etli
0.29
-
 ATP mutant R427K, pH 7.8, 30°C Rhizobium etli
0.7
-
 ATP mutant R472K, pH 7.8, 30°C Rhizobium etli
1
-
 ATP mutant R472S, pH 7.8, 30°C Rhizobium etli

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
 Rhizobium etli

Organism

Organism UniProt Comment Textmining
Rhizobium etli Q2K340
-
-
Rhizobium etli CFN 42 Q2K340
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ADP + carbamoylphosphate reverse reaction is assessed by using substrate analogue carbamoylphosphate Rhizobium etli ?
-
 r
ADP + carbamoylphosphate reverse reaction is assessed by using substrate analogue carbamoylphosphate Rhizobium etli CFN 42 ?
-
 r
ATP + pyruvate + HCO3-
-
 Rhizobium etli ADP + phosphate + oxaloacetate
-
 r
ATP + pyruvate + HCO3-
-
 Rhizobium etli CFN 42 ADP + phosphate + oxaloacetate
-
 r

Subunits

Subunits Comment Organism
tetramer
-
 Rhizobium etli

Synonyms

Synonyms Comment Organism
pyruvate carboxylase
-
 Rhizobium etli
RePC
-
 Rhizobium etli

Temperature Optimum [°C]

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

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.9
-
 ATP mutant R472K, pH 7.8, 30°C Rhizobium etli
2
-
 ATP mutant R472S, pH 7.8, 30°C Rhizobium etli
3.4
-
 ATP mutant R427K, pH 7.8, 30°C Rhizobium etli
4.2
-
 ATP mutant R427S, pH 7.8, 30°C Rhizobium etli
10.9
-
 ATP wild-type, pH 7.8, 30°C Rhizobium etli

pH Optimum

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

Cofactor

Cofactor Comment Organism Structure
biotin
-
 Rhizobium etli

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
4.5
-
 L-aspartate mutant R427K, pH 7.8, 30°C Rhizobium etli
5
-
 L-aspartate wild-type, pH 7.8, 30°C Rhizobium etli

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2
-
 ATP mutant R472S, pH 7.8, 30°C Rhizobium etli
3
-
 ATP mutant R472K, pH 7.8, 30°C Rhizobium etli
12
-
 ATP mutant R427K, pH 7.8, 30°C Rhizobium etli
26
-
 ATP mutant R427S, pH 7.8, 30°C Rhizobium etli
99
-
 ATP wild-type, pH 7.8, 30°C Rhizobium etli