Literature summary for 6.2.1.13 extracted from

Jones, C.P.; Ingram-Smith, C.
Biochemical and kinetic characterization of the recombinant ADP-forming acetyl coenzyme A synthetase from the amitochondriate protozoan Entamoeba histolytica (2014), Eukaryot. Cell, 13, 1530-1537 .

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli Rosetta2(DE3)pLysS cells	Entamoeba histolytica

Inhibitors

Inhibitors	Comment	Organism
ATP	inhibits the acetate-forming direction of the reaction	Entamoeba histolytica
Ca2+	about 3% residual activity at 20 mM	Entamoeba histolytica
Cu2+	about 15% residual activity at 20 mM	Entamoeba histolytica
D-glucose 6-phosphate	-	Entamoeba histolytica
diphosphate	inhibits the acetate-forming direction of the reaction (64% residual activity at 10 mM)	Entamoeba histolytica
glyoxylate	32% residual activity at 10 mM	Entamoeba histolytica
NAD+	47% residual activity at 10 mM	Entamoeba histolytica
Ni2+	about 3% residual activity at 20 mM	Entamoeba histolytica
Zn2+	about 5% residual activity at 20 mM	Entamoeba histolytica

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.032	-	propionyl-CoA	at pH 7.3 and 37°C	Entamoeba histolytica
0.04	-	acetyl-CoA	at pH 7.3 and 37°C	Entamoeba histolytica
0.2	-	CoA	with acetate as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
0.71	-	ADP	with propionyl-CoA as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
1.5	-	phosphate	with propionyl-CoA as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
1.6	-	ADP	with acetyl-CoA as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
1.6	-	CoA	with propionate as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
1.8	-	phosphate	with acetyl-CoA as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
1.9	-	GDP	with acetyl-CoA as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
7.2	-	ATP	with propionate as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
10	-	GTP	with acetate as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
12	-	ATP	with acetate as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
14	-	acetate	at pH 7.3 and 37°C	Entamoeba histolytica
29	-	propionate	at pH 7.3 and 37°C	Entamoeba histolytica

Metals/Ions

Metals/Ions	Comment	Organism
Co2+	90% activity in the presence of 20 mM	Entamoeba histolytica
Mg2+	100% activity in the presence of 20 mM	Entamoeba histolytica
Mn2+	100% activity in the presence of 20 mM	Entamoeba histolytica

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
150000	-	gel filtration	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	the specific activity in the acetate-forming direction is approximately half that observed in the acetyl-CoA-forming direction	ATP + acetate + CoA	-	r
ATP + acetate + CoA	Entamoeba histolytica	preferred direction of reaction	ADP + phosphate + acetyl-CoA	-	r

Organism

Organism	UniProt	Comment	Textmining
Entamoeba histolytica	C4LUV9	-	-

Purification (Commentary)

Purification (Comment)	Organism
HisTrap HP nickel affinity column chromatography, and Superdex 200 gel filtration	Entamoeba histolytica

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
ADP + phosphate + acetyl-CoA	100% activity	Entamoeba histolytica	ATP + acetate + CoA	-	r
ADP + phosphate + acetyl-CoA	the specific activity in the acetate-forming direction is approximately half that observed in the acetyl-CoA-forming direction	Entamoeba histolytica	ATP + acetate + CoA	-	r
ADP + phosphate + isobutyryl-CoA	less than 1% activity compared to acetyl-CoA	Entamoeba histolytica	ATP + isobutyrate + CoA	-	r
ADP + phosphate + isovaleryl-CoA	less than 1% activity compared to acetyl-CoA	Entamoeba histolytica	ATP + isovalerate + CoA	-	r
ADP + phosphate + propionyl-CoA	16% activity compared to acetyl-CoA	Entamoeba histolytica	ATP + propionate + CoA	-	r
ATP + acetate + CoA	100% activity	Entamoeba histolytica	ADP + phosphate + acetyl-CoA	-	r
ATP + acetate + CoA	preferred direction of reaction	Entamoeba histolytica	ADP + phosphate + acetyl-CoA	-	r
ATP + butyrate + CoA	5.6% activity compared to acetate	Entamoeba histolytica	ADP + phosphate + butyryl-CoA	-	r
ATP + heptanoate + CoA	less than 4% activity compared to acetate	Entamoeba histolytica	ADP + phosphate + heptanoyl-CoA	-	r
ATP + hexanoate + CoA	less than 4% activity compared to acetate	Entamoeba histolytica	ADP + phosphate + hexanoyl-CoA	-	r
ATP + isobutyrate + CoA	less than 4% activity compared to acetate	Entamoeba histolytica	ADP + phosphate + isobutyryl-CoA	-	r
ATP + isovalerate + CoA	less than 4% activity compared to acetate	Entamoeba histolytica	ADP + phosphate + isovaleryl-CoA	-	r
ATP + octanoate + CoA	less than 4% activity compared to acetate	Entamoeba histolytica	ADP + phosphate + octanoyl-CoA	-	r
ATP + phenylacetate + CoA	less than 4% activity compared to acetate	Entamoeba histolytica	ADP + phosphate + phenylacetyl-CoA	-	r
ATP + propionate + CoA	72% activity compared to acetate	Entamoeba histolytica	ADP + phosphate + propionyl-CoA	-	r
ATP + succinate + CoA	less than 4% activity compared to acetate	Entamoeba histolytica	ADP + phosphate + succinyl-CoA	-	r
ATP + valerate + CoA	less than 4% activity compared to acetate	Entamoeba histolytica	ADP + phosphate + valeryl-CoA	-	r
CDP + phosphate + acetyl-CoA	7% activity compared to ADP	Entamoeba histolytica	CTP + acetate + CoA	-	r
CTP + acetate + CoA	4.1% activity compared to ATP	Entamoeba histolytica	CDP + phosphate + acetyl-CoA	-	r
GDP + phosphate + acetyl-CoA	106% activity compared to ADP	Entamoeba histolytica	GTP + acetate + CoA	-	r
GTP + acetate + CoA	57% activity compared to ATP	Entamoeba histolytica	GDP + phosphate + acetyl-CoA	-	r
IDP + phosphate + acetyl-CoA	28% activity compared to ADP	Entamoeba histolytica	ITP + acetate + CoA	-	r
ITP + acetate + CoA	15% activity compared to ATP	Entamoeba histolytica	IDP + phosphate + acetyl-CoA	-	r
additional information	no activity with diphosphate, AMP, TDP, TTP, butyryl-CoA, succinyl-CoA, or phenylacetyl-CoA	Entamoeba histolytica	?	-	?
UDP + phosphate + acetyl-CoA	2.8% activity compared to ADP	Entamoeba histolytica	UTP + acetate + CoA	-	r
UTP + acetate + CoA	1.6% activity compared to ATP	Entamoeba histolytica	UDP + phosphate + acetyl-CoA	-	r

Subunits

Subunits	Comment	Organism
homodimer	2 * 78999, calculated from amino acid sequence	Entamoeba histolytica

Synonyms

Synonyms	Comment	Organism
ACD	-	Entamoeba histolytica
acetate:CoA ligase [ADP-forming]	-	Entamoeba histolytica
ADP-forming acetyl-coenzyme A synthetase	-	Entamoeba histolytica

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
55	-	-	Entamoeba histolytica

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
37	-	the enzyme exhibits 70% activity at 37°C	Entamoeba histolytica

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
18	-	phosphate	with propionyl-CoA as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
21	-	propionyl-CoA	at pH 7.3 and 37°C	Entamoeba histolytica
24	-	ADP	with propionyl-CoA as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
110	-	acetyl-CoA	at pH 7.3 and 37°C	Entamoeba histolytica
120	-	phosphate	with acetyl-CoA as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
140	-	ADP	with acetyl-CoA as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
170	-	GDP	with acetyl-CoA as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
180	-	GTP	with acetate as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
190	-	propionate	at pH 7.3 and 37°C	Entamoeba histolytica
220	-	CoA	with acetate as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
240	-	acetate	at pH 7.3 and 37°C	Entamoeba histolytica
240	-	ATP	with propionate as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
260	-	CoA	with propionate as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
320	-	ATP	with acetate as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica

Cofactor

Cofactor	Comment	Organism
ADP	-	Entamoeba histolytica
ATP	-	Entamoeba histolytica
GDP	-	Entamoeba histolytica
GTP	-	Entamoeba histolytica

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor
0.81	-	at pH 7.3 and 37°C	Entamoeba histolytica	ATP
0.83	-	at pH 7.3 and 37°C	Entamoeba histolytica	diphosphate
10	-	at pH 7.3 and 37°C	Entamoeba histolytica	D-glucose 6-phosphate

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
6.3	-	propionate	at pH 7.3 and 37°C	Entamoeba histolytica
12	-	phosphate	with propionyl-CoA as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
16	-	acetate	at pH 7.3 and 37°C	Entamoeba histolytica
18	-	GTP	with acetate as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
27	-	ATP	with acetate as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
33	-	ATP	with propionate as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
34	-	ADP	with propionyl-CoA as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
65	-	phosphate	with acetyl-CoA as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
89	-	ADP	with acetyl-CoA as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
90	-	GDP	with acetyl-CoA as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
160	-	CoA	with propionate as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
650	-	propionyl-CoA	at pH 7.3 and 37°C	Entamoeba histolytica
1100	-	CoA	with acetate as cosubstrate, at pH 7.3 and 37°C	Entamoeba histolytica
2600	-	acetyl-CoA	at pH 7.3 and 37°C	Entamoeba histolytica