Cloned (Comment) | Organism |
---|---|
recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli strain Rosetta2(DE3)pLysS | Entamoeba histolytica |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
acetyl phosphate | - |
Entamoeba histolytica | |
AMP | slight inhibition | Entamoeba histolytica | |
ATP | ATP inhibits the acetate-forming direction of the reaction, with 50% inhibitory concentration of 0.81 mM, which is in the range of its physiological concentration. Diphosphate displays mixed inhibition versus each of the three substrates, acetyl-CoA, ADP, and phosphate. No inhibition by lactate and pyruvate | Entamoeba histolytica | |
cAMP | - |
Entamoeba histolytica | |
D-fructose 1,6-bisphosphate | - |
Entamoeba histolytica | |
D-fructose 6-phosphate | - |
Entamoeba histolytica | |
D-glucose 6-phosphate | - |
Entamoeba histolytica | |
diphosphate | diphosphate inhibits the acetate-forming direction of the reaction, with 50% inhibitory concentration of 0.75 mM, which is in the range of its physiological concentration. ATP displays mixed inhibition versus each of the three substrates, acetyl-CoA, ADP, and phosphate | Entamoeba histolytica | |
glyoxylate | inhibits the activity in the acetate-forming direction | Entamoeba histolytica | |
NAD+ | - |
Entamoeba histolytica | |
NADH | - |
Entamoeba histolytica |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten steady-state kinetic analysis, pseudo-first-order reaction kinetic determinations are performed in both directions of the reaction | Entamoeba histolytica | |
0.032 | - |
propionyl-CoA | pH 7.3, 37°C, propionate-forming direction, with propionyl-CoA | Entamoeba histolytica | |
0.04 | - |
acetyl-CoA | pH 7.3, 37°C, acetate-forming direction | Entamoeba histolytica | |
0.2 | - |
CoA | pH 7.3, 37°C, acetyl-CoA-forming direction, with acetate | Entamoeba histolytica | |
0.71 | - |
ADP | pH 7.3, 37°C, propionate-forming direction, with propionyl-CoA | Entamoeba histolytica | |
1.5 | - |
phosphate | pH 7.3, 37°C, propionate-forming direction, with propionyl-CoA | Entamoeba histolytica | |
1.6 | - |
ADP | pH 7.3, 37°C, acetate-forming direction, with acetyl-CoA | Entamoeba histolytica | |
1.6 | - |
CoA | pH 7.3, 37°C, propionyl-CoA-forming direction, with propionate | Entamoeba histolytica | |
1.8 | - |
phosphate | pH 7.3, 37°C, acetate-forming direction, with acetyl-CoA | Entamoeba histolytica | |
1.9 | - |
GDP | pH 7.3, 37°C, acetate-forming direction, with acetyl-CoA | Entamoeba histolytica | |
7.2 | - |
ATP | pH 7.3, 37°C, propionyl-CoA-forming direction, with propionate | Entamoeba histolytica | |
10 | - |
GTP | pH 7.3, 37°C, acetyl-CoA-forming direction, with acetate | Entamoeba histolytica | |
12 | - |
ATP | pH 7.3, 37°C, acetyl-CoA-forming direction, with acetate | Entamoeba histolytica | |
14 | - |
acetate | pH 7.3, 37°C, acetyl-CoA-forming direction | Entamoeba histolytica | |
29 | - |
propionate | pH 7.3, 37°C, propionyl-CoA-forming direction, with propionate | Entamoeba histolytica |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | can slightly substitute for Mg2+ | Entamoeba histolytica | |
Co2+ | can partially substitute for Mg2+ | Entamoeba histolytica | |
Cu2+ | can slightly substitute for Mg2+ | Entamoeba histolytica | |
Mg2+ | required for activity, best divalent cation | Entamoeba histolytica | |
Mn2+ | can substitute for Mg2+ | Entamoeba histolytica | |
Ni2+ | can slightly substitute for Mg2+ | Entamoeba histolytica | |
Zn2+ | can slightly substitute for Mg2+ | Entamoeba histolytica |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
150000 | - |
recombinant His6-tagged enzyme, gel filtration | Entamoeba histolytica |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + acetate + CoA | Entamoeba histolytica | - |
ADP + phosphate + acetyl-CoA | - |
r | |
GTP + acetate + CoA | Entamoeba histolytica | - |
GDP + phosphate + acetyl-CoA | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Entamoeba histolytica | Q9NAT4 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged enzyme from Escherichia coli strain Rosetta2(DE3)pLysS by nickel affinity chromatography | Entamoeba histolytica |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
trophozoite | - |
Entamoeba histolytica | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ADP + phosphate + acetyl-CoA | - |
Entamoeba histolytica | ATP + acetate + CoA | - |
r | |
ADP + phosphate + propionyl-CoA | - |
Entamoeba histolytica | ATP + propionate + CoA | - |
r | |
ATP + acetate + CoA | - |
Entamoeba histolytica | ADP + phosphate + acetyl-CoA | - |
r | |
ATP + propionate + CoA | - |
Entamoeba histolytica | ADP + phosphate + propionyl-CoA | - |
r | |
GDP + phosphate + acetyl-CoA | - |
Entamoeba histolytica | GTP + acetate + CoA | - |
r | |
GTP + acetate + CoA | - |
Entamoeba histolytica | GDP + phosphate + acetyl-CoA | - |
r | |
GTP + acetate + CoA | GTP gives 57% of the activity with ATP | Entamoeba histolytica | GDP + phosphate + acetyl-CoA | - |
r | |
ITP + acetate + CoA | ITP gives 15% of the activity with ATP | Entamoeba histolytica | IDP + phosphate + acetyl-CoA | - |
r | |
additional information | the enzyme can utilize both ADP/ATP and GDP/GTP in the respective directions of the reaction. Less than 4% of maximal activity is observed with isobutyrate, valerate, isovalerate, hexanoate, heptanoate, octanoate, succinate, or phenylacetate as the acyl substrate. Activity observed with ATP (representing 100% activity) is nearly double that with GTP (57%) and substantially higher than the activities observed with ITP (15%), CTP (4.1%), or UTP (1.6%), no activity is observed with TTP or diphosphate. Enzymatic activity in the acetate-forming direction is determined by measuring the release of CoA with a sulfhydryl group (CoASH) from acyl-CoA by using Ellman's thiol reagent (DTNB). Production of NTB2- by CoASH cleavage of DTNB is measured spectrophotometrically at 412 nm. Activity in the acetate-forming direction is confirmed using the hexokinase/glucose-6-phosphate dehydrogenase-coupled assay. This assay couples ATP formation to the reduction of NADP+ to NADPH, which is measured spectrophotometrically at 340 nm | Entamoeba histolytica | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 78999, recombinant His6-tagged enzyme, sequence calculation and SDS-PAGE | Entamoeba histolytica |
Synonyms | Comment | Organism |
---|---|---|
ACD | - |
Entamoeba histolytica |
Acetyl-CoA synthetase (ADP-forming) | - |
Entamoeba histolytica |
ADP-forming acetyl coenzyme A synthetase | - |
Entamoeba histolytica |
EhACD | - |
Entamoeba histolytica |
More | cf. EC 6.2.1.13 | Entamoeba histolytica |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
acetate-forming assay at | Entamoeba histolytica |
55 | - |
acetyl-CoA-forming direction | Entamoeba histolytica |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
18 | - |
phosphate | pH 7.3, 37°C, acetate-forming direction, with propionyl-CoA | Entamoeba histolytica | |
21 | - |
propionyl-CoA | pH 7.3, 37°C, acetate-forming direction, with propionyl-CoA | Entamoeba histolytica | |
24 | - |
ADP | pH 7.3, 37°C, acetate-forming direction, with propionyl-CoA | Entamoeba histolytica | |
110 | - |
acetyl-CoA | pH 7.3, 37°C, acetate-forming direction | Entamoeba histolytica | |
120 | - |
phosphate | pH 7.3, 37°C, acetate-forming direction, with acetyl-CoA | Entamoeba histolytica | |
140 | - |
ADP | pH 7.3, 37°C, acetate-forming direction, with acetyl-CoA | Entamoeba histolytica | |
170 | - |
GDP | pH 7.3, 37°C, acetate-forming direction, with acetyl-CoA | Entamoeba histolytica | |
180 | - |
GTP | pH 7.3, 37°C, acetyl-CoA-forming direction, with acetate | Entamoeba histolytica | |
190 | - |
propionate | pH 7.3, 37°C, propionyl-CoA-forming direction, with propionate | Entamoeba histolytica | |
220 | - |
CoA | pH 7.3, 37°C, acetyl-CoA-forming direction, with acetate | Entamoeba histolytica | |
240 | - |
acetate | pH 7.3, 37°C, acetyl-CoA-forming direction | Entamoeba histolytica | |
240 | - |
ATP | pH 7.3, 37°C, propionyl-CoA-forming direction, with propionate | Entamoeba histolytica | |
260 | - |
CoA | pH 7.3, 37°C, propionyl-CoA-forming direction, with propionate | Entamoeba histolytica | |
320 | - |
ATP | pH 7.3, 37°C, acetyl-CoA-forming direction, with acetate | Entamoeba histolytica |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.3 | - |
assay at | Entamoeba histolytica |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ADP | - |
Entamoeba histolytica | |
ATP | - |
Entamoeba histolytica | |
GDP | - |
Entamoeba histolytica | |
GTP | - |
Entamoeba histolytica |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.81 | - |
pH 7.3, 37°C, acetate-forming direction | Entamoeba histolytica | ATP | |
0.83 | - |
pH 7.3, 37°C, acetate-forming direction | Entamoeba histolytica | diphosphate |
General Information | Comment | Organism |
---|---|---|
physiological function | enzyme ACD is regulated through inhibition by ATP and diphosphate in the acetate-forming direction | Entamoeba histolytica |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
6.3 | - |
propionate | pH 7.3, 37°C, propionyl-CoA-forming direction, with propionate | Entamoeba histolytica | |
12 | - |
phosphate | pH 7.3, 37°C, acetate-forming direction, with propionyl-CoA | Entamoeba histolytica | |
16 | - |
acetate | pH 7.3, 37°C, acetyl-CoA-forming direction | Entamoeba histolytica | |
18 | - |
GTP | pH 7.3, 37°C, acetyl-CoA-forming direction, with acetate | Entamoeba histolytica | |
27 | - |
ATP | pH 7.3, 37°C, acetyl-CoA-forming direction, with acetate | Entamoeba histolytica | |
33 | - |
ATP | pH 7.3, 37°C, propionyl-CoA-forming direction, with propionate | Entamoeba histolytica | |
34 | - |
ADP | pH 7.3, 37°C, acetate-forming direction, with propionyl-CoA | Entamoeba histolytica | |
65 | - |
phosphate | pH 7.3, 37°C, acetate-forming direction, with acetyl-CoA | Entamoeba histolytica | |
89 | - |
ADP | pH 7.3, 37°C, acetate-forming direction, with acetyl-CoA | Entamoeba histolytica | |
90 | - |
GDP | pH 7.3, 37°C, acetate-forming direction, with acetyl-CoA | Entamoeba histolytica | |
160 | - |
CoA | pH 7.3, 37°C, propionyl-CoA-forming direction, with propionate | Entamoeba histolytica | |
650 | - |
propionyl-CoA | pH 7.3, 37°C, acetate-forming direction, with propionyl-CoA | Entamoeba histolytica | |
1100 | - |
CoA | pH 7.3, 37°C, acetyl-CoA-forming direction, with acetate | Entamoeba histolytica | |
2600 | - |
acetyl-CoA | pH 7.3, 37°C, acetate-forming direction | Entamoeba histolytica |