Cloned (Comment) | Organism |
---|---|
genes acdIa and acdIb encoding the alpha and beta subunits, expression in Escherichia coli strain BL21(DE3) | Pyrococcus furiosus |
Protein Variants | Comment | Organism |
---|---|---|
D212Ebeta | site-directed mutagenesis, the mutant shows 2-4% of wild-type activity, slightly impaired in arsenolysis | Pyrococcus furiosus |
D212Nbeta | site-directed mutagenesis, the mutant shows highly reduced phosphorylation/phosphorolysis activity, but only slightly impaired in arsenolysis | Pyrococcus furiosus |
E218Qalpha | site-directed mutagenesis, inactive mutant | Pyrococcus furiosus |
H257Dalpha | site-directed mutagenesis, inactive mutant | Pyrococcus furiosus |
H71Abeta | site-directed mutagenesis, inactive mutant concerning phosphorylation/phosphorolysis, slightly impaired in arsenolysis | Pyrococcus furiosus |
additional information | all mutations in the alpha-subunit have dramatic effects on arsenolysis activity, while mutations in the beta-subunit cause only moderate loss of activity | Pyrococcus furiosus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetic parameters of wild type and mutant enzymes at 55°C, overview | Pyrococcus furiosus | |
0.0037 | - |
acetyl-CoA | pH 7.5, 80°C, wild-type alpha-subunit | Pyrococcus furiosus | |
0.0039 | - |
acetyl-CoA | pH 7.5, 55°C, wild-type enzyme | Pyrococcus furiosus | |
0.0042 | - |
acetyl-CoA | pH 7.5, 80°C, wild-type enzyme | Pyrococcus furiosus | |
0.0139 | - |
CoA | pH 7.5, 55°C, wild-type enzyme | Pyrococcus furiosus | |
0.093 | - |
ADP | pH 7.5, 55°C, wild-type enzyme | Pyrococcus furiosus | |
0.221 | - |
ATP | pH 7.5, 55°C, wild-type enzyme | Pyrococcus furiosus | |
0.23 | - |
phosphate | pH 7.5, 80°C, wild-type enzyme | Pyrococcus furiosus | |
0.272 | - |
phosphate | pH 7.5, 55°C, wild-type enzyme | Pyrococcus furiosus | |
0.625 | - |
acetate | pH 7.5, 55°C, wild-type enzyme | Pyrococcus furiosus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Pyrococcus furiosus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + acetate + CoA | Pyrococcus furiosus | - |
AMP + diphosphate + acetyl-CoA | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pyrococcus furiosus | - |
genes acdIa and acdIb encoding the alpha and beta subunits | - |
Purification (Comment) | Organism |
---|---|
recombinant alpha and beta subunits from Escherichia coli strain BL21(DE3) by heat tteatment at 90°C for 30 min, hydrophobic interaction chromatography, and gel filtration | Pyrococcus furiosus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
ATP + acetate + CoA = AMP + diphosphate + acetyl-CoA | the ACD reaction follows a four-step mechanism including transient phosphorylation of two active site histidine residues, structure and reaction mechanism of ACD | Pyrococcus furiosus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ADP + phosphate + acetyl-CoA | - |
Pyrococcus furiosus | ATP + acetate + CoA | - |
? | |
ATP + acetate + CoA | - |
Pyrococcus furiosus | AMP + diphosphate + acetyl-CoA | - |
? | |
ATP + acetate + CoA | formation of enzyme-bound acetyl phosphate and enzyme phosphorylation at His257alpha, respectively. The phosphoryl group is transferred from the His257alpha to ADP via transient phosphorylation of a second conserved histidine residue in the beta-subunit, His71beta | Pyrococcus furiosus | AMP + diphosphate + acetyl-CoA | - |
ir | |
additional information | the enzyme performs arsenolysis, the alpha-subunit alone also catalyzes arsenolysis, overview | Pyrococcus furiosus | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | structural analysis by circular dichroism spectroscopy and structure modelling, comparison of domain organization of subunits to succinyl-CoA synthetases, EC 6.2.1.5, overview | Pyrococcus furiosus |
Synonyms | Comment | Organism |
---|---|---|
ACD | - |
Pyrococcus furiosus |
ADP-forming acetyl-CoA synthetase | - |
Pyrococcus furiosus |
More | ACD belongs to the protein superfamily of nucleoside diphosphate-forming acyl-CoA synthetases | Pyrococcus furiosus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
55 | - |
phosphorylation and phosphorolysis assays at | Pyrococcus furiosus |
80 | - |
arsenolytic assay at and temperature optimum | Pyrococcus furiosus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Pyrococcus furiosus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ADP | - |
Pyrococcus furiosus | |
ATP | - |
Pyrococcus furiosus |