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Literature summary for 6.2.1.13 extracted from
- Investigating the mechanism of ADP-forming acetyl-CoA synthetase from the protozoan parasite Entamoeba histolytica (2017), FEBS Lett., 591, 603-612 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli Rosetta2(DE3) pLysS cells | Entamoeba histolytica |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D674A | site-directed mutagenesis, the mutation does not eliminate the ability of ACD to be phosphorylated from either direction, although phosphorylation by ATP is reduced | Entamoeba histolytica |
| D674A | the variant retains about 1% activity compared to wild type enzyme | Entamoeba histolytica |
| D674E | the variant retains about 45% activity compared to wild type enzyme | Entamoeba histolytica |
| D674N | the variant retains about 1% activity compared to wild type enzyme | Entamoeba histolytica |
| E213A | site-directed mutagenesis, the mutant variant is not phosphorylated by acetyl-CoA and phosphate | Entamoeba histolytica |
| E213A | the variant retains about 1% activity compared to wild type enzyme | Entamoeba histolytica |
| E213D | the variant retains about 1% activity compared to wild type enzyme | Entamoeba histolytica |
| E213Q | the variant retains about 1% activity compared to wild type enzyme | Entamoeba histolytica |
| E218A | site-directed mutagenesis, the mutant variant is not phosphorylated by acetyl-CoA and phosphate | Entamoeba histolytica |
| H252A | site-directed mutagenesis, alteration of His252 in EhACD effectively eliminate overall enzymatic activity as well as phosphorylation in either direction of the reaction | Entamoeba histolytica |
| H252A | the variant has no detectable activity in either direction of the reaction | Entamoeba histolytica |
| H533D | the variant retains about 3% activity compared to wild type enzyme | Entamoeba histolytica |
| H533E | the variant retains about 1% activity compared to wild type enzyme | Entamoeba histolytica |
| H533K | the variant retains about 10% activity compared to wild type enzyme | Entamoeba histolytica |
| H533N | the variant retains about 5% activity compared to wild type enzyme | Entamoeba histolytica |
| H533Q | the variant retains about 5% activity compared to wild type enzyme | Entamoeba histolytica |
| H533R | the variant retains about 10% activity compared to wild type enzyme | Entamoeba histolytica |
| H533R | the variant retains less than 25% activity compared to wild type enzyme | Entamoeba histolytica |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.01 | - |
acetyl-CoA | mutant enzyme H533A, pH and temperature not specified in the publication | Entamoeba histolytica |  |
| 0.011 | - |
CoA | mutant enzyme H533D, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.011 | - |
CoA | mutant enzyme H533Q, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.012 | - |
acetyl-CoA | mutant enzyme H533E, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.015 | - |
acetyl-CoA | mutant enzyme H533N, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.016 | - |
acetyl-CoA | mutant enzyme H533R, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.019 | - |
acetyl-CoA | mutant enzyme H533Q, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.027 | - |
acetyl-CoA | mutant enzyme H533D, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.03 | - |
acetyl-CoA | mutant enzyme H533K, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.043 | - |
acetyl-CoA | wild type enzyme, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.07 | - |
CoA | mutant enzyme H533A, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.08 | - |
CoA | mutant enzyme H533N, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.1 | - |
CoA | mutant enzyme H533R, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.13 | - |
CoA | mutant enzyme H533K, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.15 | - |
phosphate | mutant enzyme H533R, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.18 | - |
phosphate | mutant enzyme H533E, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.2 | - |
CoA | wild type enzyme, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.25 | - |
phosphate | mutant enzyme H533D, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.26 | - |
CoA | mutant enzyme H533E, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.31 | - |
acetate | mutant enzyme H533D, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.4 | - |
acetate | mutant enzyme H533E, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.41 | - |
ADP | mutant enzyme H533A, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.55 | - |
acetate | mutant enzyme H533Q, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.57 | - |
ADP | mutant enzyme H533N, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.6 | - |
ADP | mutant enzyme H533D, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.64 | - |
ADP | mutant enzyme H533R, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.68 | - |
acetate | mutant enzyme H533A, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.75 | - |
phosphate | mutant enzyme H533A, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.76 | - |
acetate | mutant enzyme H533N, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.84 | - |
acetate | mutant enzyme H533K, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.86 | - |
phosphate | mutant enzyme H533Q, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.94 | - |
phosphate | mutant enzyme H533N, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.97 | - |
ADP | mutant enzyme H533K, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.19 | - |
ADP | mutant enzyme H533E, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.2 | - |
ATP | mutant enzyme H533D, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.28 | - |
ADP | mutant enzyme H533Q, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.56 | - |
ADP | wild type enzyme, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.7 | - |
ATP | mutant enzyme H533K, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.7 | - |
acetate | mutant enzyme H533R, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.77 | - |
phosphate | wild type enzyme, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 2.2 | - |
ATP | mutant enzyme H533A, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 2.5 | - |
ATP | mutant enzyme H533Q, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 2.5 | - |
ATP | mutant enzyme H533R, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 3.25 | - |
phosphate | mutant enzyme H533K, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 3.9 | - |
ATP | mutant enzyme H533E, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 7.7 | - |
ATP | mutant enzyme H533N, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 12 | - |
ATP | wild type enzyme, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 14 | - |
acetate | wild type enzyme, pH and temperature not specified in the publication | Entamoeba histolytica | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | required | Entamoeba histolytica | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ADP + phosphate + acetyl-CoA | Entamoeba histolytica | - |
ATP + acetate + CoA | - |
r | |
| ATP + acetate + CoA | Entamoeba histolytica | - |
ADP + phosphate + acetyl-CoA | - |
r | |
| additional information | Entamoeba histolytica | ADP-forming acetyl-CoA synthetase (ACD) catalyzes the interconversion of acetyl-CoA and acetate | ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Entamoeba histolytica | C4LUV9 | - |
- |
| Entamoeba histolytica | Q9NAT4 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| nickel affinity column chromatography | Entamoeba histolytica |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| ATP + acetate + CoA = ADP + phosphate + acetyl-CoA | the enzyme uses a four-step reaction mechanism including an additional phosphoryl transfer step, the important His252, located in the beta subunit, is invovled | Entamoeba histolytica |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ADP + phosphate + acetyl-CoA | - |
Entamoeba histolytica | ATP + acetate + CoA | - |
r | |
| ATP + acetate + CoA | - |
Entamoeba histolytica | ADP + phosphate + acetyl-CoA | - |
r | |
| additional information | ADP-forming acetyl-CoA synthetase (ACD) catalyzes the interconversion of acetyl-CoA and acetate | Entamoeba histolytica | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| More | ACD subunit structure , overview | Entamoeba histolytica |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| ACD | - |
Entamoeba histolytica |
| ADP-forming acetyl-CoA synthetase | - |
Entamoeba histolytica |
| EhACD | - |
Entamoeba histolytica |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.12 | - |
acetyl-CoA | mutant enzyme H533E, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.18 | - |
acetate | mutant enzyme H533D, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.22 | - |
phosphate | mutant enzyme H533E, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.25 | - |
ATP | mutant enzyme H533D, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.25 | - |
CoA | mutant enzyme H533D, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.26 | - |
acetate | mutant enzyme H533E, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.27 | - |
ADP | mutant enzyme H533E, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.27 | - |
CoA | mutant enzyme H533E, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.28 | - |
ATP | mutant enzyme H533E, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.2 | - |
acetate | mutant enzyme H533Q, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.3 | - |
CoA | mutant enzyme H533Q, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.4 | - |
ADP | mutant enzyme H533N, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.4 | - |
acetyl-CoA | mutant enzyme H533N, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.4 | - |
ATP | mutant enzyme H533Q, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.5 | - |
phosphate | mutant enzyme H533N, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.5 | - |
acetate | mutant enzyme H533N, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.6 | - |
acetyl-CoA | mutant enzyme H533D, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.7 | - |
ADP | mutant enzyme H533D, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.7 | - |
phosphate | mutant enzyme H533D, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.7 | - |
acetyl-CoA | mutant enzyme H533Q, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.8 | - |
phosphate | mutant enzyme H533Q, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 2 | - |
ATP | mutant enzyme H533N, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 2 | - |
CoA | mutant enzyme H533N, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 2.3 | - |
phosphate | mutant enzyme H533A, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 2.4 | - |
acetyl-CoA | mutant enzyme H533K, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 2.5 | - |
ADP | mutant enzyme H533Q, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 2.7 | - |
acetyl-CoA | mutant enzyme H533A, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 2.8 | - |
ADP | mutant enzyme H533A, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 3.1 | - |
phosphate | mutant enzyme H533K, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 3.3 | - |
ATP | mutant enzyme H533A, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 3.3 | - |
acetate | mutant enzyme H533A, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 3.3 | - |
ADP | mutant enzyme H533K, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 3.4 | - |
acetate | mutant enzyme H533K, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 3.7 | - |
ATP | mutant enzyme H533K, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 4.1 | - |
CoA | mutant enzyme H533K, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 4.2 | - |
CoA | mutant enzyme H533A, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 5.6 | - |
acetate | mutant enzyme H533R, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 6.6 | - |
ATP | mutant enzyme H533R, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 7 | - |
CoA | mutant enzyme H533R, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 14 | - |
phosphate | mutant enzyme H533R, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 18 | - |
acetyl-CoA | mutant enzyme H533R, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 20 | - |
ADP | mutant enzyme H533R, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 111 | - |
acetyl-CoA | wild type enzyme, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 115 | - |
phosphate | wild type enzyme, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 138 | - |
ADP | wild type enzyme, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 233 | - |
acetate | wild type enzyme, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 320 | - |
ATP | wild type enzyme, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 328 | - |
CoA | wild type enzyme, pH and temperature not specified in the publication | Entamoeba histolytica | |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| ADP | - |
Entamoeba histolytica | |
| ATP | - |
Entamoeba histolytica | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | the enzyme is a member of the NDP-forming acyl-CoA synthetase enzyme superfamily | Entamoeba histolytica |
| additional information | residue His252 is essential for enzymatic activity, substrate binding structure and mechanism, overview | Entamoeba histolytica |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.06 | - |
acetyl-CoA | mutant enzyme H533D, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.07 | - |
ATP | mutant enzyme H533E, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.09 | - |
acetyl-CoA | mutant enzyme H533Q, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.2 | - |
ADP | mutant enzyme H533E, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.21 | - |
ATP | mutant enzyme H533D, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.26 | - |
ATP | mutant enzyme H533N, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.57 | - |
CoA | mutant enzyme H533Q, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.59 | - |
acetate | mutant enzyme H533D, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.67 | - |
acetate | mutant enzyme H533E, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 0.97 | - |
phosphate | mutant enzyme H533K, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.1 | - |
CoA | mutant enzyme H533E, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.3 | - |
phosphate | mutant enzyme H533E, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.6 | - |
ATP | mutant enzyme H533A, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.7 | - |
phosphate | mutant enzyme H533N, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.9 | - |
acetate | mutant enzyme H533N, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1.9 | - |
ADP | mutant enzyme H533Q, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 2.1 | - |
phosphate | mutant enzyme H533Q, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 2.2 | - |
acetate | mutant enzyme H533Q, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 2.4 | - |
ATP | mutant enzyme H533K, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 2.5 | - |
ADP | mutant enzyme H533N, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 2.7 | - |
ATP | mutant enzyme H533R, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 2.9 | - |
ADP | mutant enzyme H533D, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 3.1 | - |
phosphate | mutant enzyme H533A, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 3.3 | - |
acetate | mutant enzyme H533R, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 3.4 | - |
ADP | mutant enzyme H533K, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 4.1 | - |
acetate | mutant enzyme H533K, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 4.8 | - |
acetate | mutant enzyme H533A, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 6.9 | - |
ADP | mutant enzyme H533A, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 7.3 | - |
phosphate | mutant enzyme H533D, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 10 | - |
acetyl-CoA | mutant enzyme H533E, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 16 | - |
acetate | wild type enzyme, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 23 | - |
CoA | mutant enzyme H533D, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 26 | - |
CoA | mutant enzyme H533N, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 27 | - |
ATP | wild type enzyme, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 32 | - |
ADP | mutant enzyme H533R, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 60 | - |
CoA | mutant enzyme H533A, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 65 | - |
phosphate | wild type enzyme, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 70 | - |
CoA | mutant enzyme H533R, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 79 | - |
acetyl-CoA | mutant enzyme H533K, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 89 | - |
ADP | wild type enzyme, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 92 | - |
acetyl-CoA | mutant enzyme H533N, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 95 | - |
phosphate | mutant enzyme H533R, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 116 | - |
ATP | mutant enzyme H533Q, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 266 | - |
acetyl-CoA | mutant enzyme H533A, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 287 | - |
CoA | mutant enzyme H533K, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1100 | - |
CoA | wild type enzyme, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 1115 | - |
acetyl-CoA | mutant enzyme H533R, pH and temperature not specified in the publication | Entamoeba histolytica | |
| 2600 | - |
acetyl-CoA | wild type enzyme, pH and temperature not specified in the publication | Entamoeba histolytica | |
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