Literature summary for 2.3.1.8 extracted from

Lawrence, S.H.; Luther, K.B.; Schindelin, H.; Ferry, J.G.
Structural and functional studies suggest a catalytic mechanism for the phosphotransacetylase from Methanosarcina thermophila (2006), J. Bacteriol., 188, 1143-1154.

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli	Methanosarcina thermophila

Crystallization (Commentary)

Crystallization (Comment)	Organism
hanging drop vapor diffusion method. Crystal structures of phosphotransacetylase in complex with the substrate CoA reveals one CoA (CoA(1)) bound in the proposed active site cleft and an additional CoA (CoA(2)) bound at the periphery of the cleft. The crystal structures indicat that binding of CoA(1) is mediated by a series of hydrogen bonds and extensive van der Waals interactions with the enzyme and that there are fewer of these interactions between CoA(2) and the enzyme	Methanosarcina thermophila

Crystallization (Comment)

Organism

hanging drop vapor diffusion method. Crystal structures of phosphotransacetylase in complex with the substrate CoA reveals one CoA (CoA(1)) bound in the proposed active site cleft and an additional CoA (CoA(2)) bound at the periphery of the cleft. The crystal structures indicat that binding of CoA(1) is mediated by a series of hydrogen bonds and extensive van der Waals interactions with the enzyme and that there are fewer of these interactions between CoA(2) and the enzyme

Methanosarcina thermophila

Protein Variants

Protein Variants	Comment	Organism
D316E	kcat for the reaction of acetyl phosphate and CoA is 2.4fold lower than wild-type value, Km for CoA is 1.1fold higher than wild-type value, Km for acetyl phosphate is 1.4fold lower than wild-type value	Methanosarcina thermophila
R310A	kcat for the reaction of acetyl phosphate and CoA is 22.6fold lower than wild-type value, Km for CoA is 1.8fold higher than wild-type value, Km for acetyl phosphate is 122fold higher than wild-type value	Methanosarcina thermophila
R310K	kcat for the reaction of acetyl phosphate and CoA is 472fold lower than wild-type value, Km for CoA is 1.8fold higher than wild-type value, Km for acetyl phosphate is 2.9fold higher than wild-type value	Methanosarcina thermophila
R310Q	kcat for the reaction of acetyl phosphate and CoA is 75.2fold lower than wild-type value, Km for CoA is 2.8fold higher than wild-type value, Km for acetyl phosphate is 4.2fold higher than wild-type value	Methanosarcina thermophila
S309A	kcat for the reaction of acetyl phosphate and CoA is 358fold lower than wild-type value, Km for CoA is nearly identical to wild-type value, Km for acetyl phosphate is 1.96fold lower than wild-type value	Methanosarcina thermophila
S309C	kcat for the reaction of acetyl phosphate and CoA is 851fold lower than wild-type value, Km for CoA is1.4 fold higher than wild-type value, Km for acetyl phosphate is 1.4fold higher than wild-type value	Methanosarcina thermophila
S309T	kcat for the reaction of acetyl phosphate and CoA is 337fold lower than wild-type value, Km for CoA is 1.8fold lower than wild-type value, Km for acetyl phosphate is nearly identical to wild-type value	Methanosarcina thermophila

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.037	-	CoA	23°C, pH 7.2, mutant enzyme S309T	Methanosarcina thermophila
0.065	-	CoA	23°C, pH 7.2, wild-type enzyme	Methanosarcina thermophila
0.067	-	CoA	23°C, pH 7.2, mutant enzyme S309A	Methanosarcina thermophila
0.074	-	CoA	23°C, pH 7.2, mutant enzyme D316E	Methanosarcina thermophila
0.094	-	acetyl phosphate	23°C, pH 7.2, mutant enzyme S309A	Methanosarcina thermophila
0.094	-	CoA	23°C, pH 7.2, mutant enzyme S309C	Methanosarcina thermophila
0.116	-	CoA	23°C, pH 7.2, mutant enzyme R310K	Methanosarcina thermophila
0.12	-	CoA	23°C, pH 7.2, mutant enzyme R310A	Methanosarcina thermophila
0.143	-	acetyl phosphate	23°C, pH 7.2, mutant enzyme D316E	Methanosarcina thermophila
0.175	-	acetyl phosphate	23°C, pH 7.2, mutant enzyme S309T	Methanosarcina thermophila
0.185	-	CoA	23°C, pH 7.2, mutant enzyme R310Q	Methanosarcina thermophila
0.185	-	acetyl phosphate	23°C, pH 7.2, wild-type enzyme	Methanosarcina thermophila
0.255	-	acetyl phosphate	23°C, pH 7.2, mutant enzyme S309C	Methanosarcina thermophila
0.531	-	acetyl phosphate	23°C, pH 7.2, mutant enzyme R310K	Methanosarcina thermophila
0.775	-	acetyl phosphate	23°C, pH 7.2, mutant enzyme R310Q	Methanosarcina thermophila
22.5	-	acetyl phosphate	23°C, pH 7.2, mutant enzyme R310A	Methanosarcina thermophila

Organism

Organism	UniProt	Comment	Textmining
Methanosarcina thermophila	P38503	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Methanosarcina thermophila

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
acetyl-phosphate + CoA	-	Methanosarcina thermophila	acetyl-CoA + phosphate	-	?

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
6.1	-	CoA	23°C, pH 7.2, mutant enzyme S309C	Methanosarcina thermophila
11	-	CoA	23°C, pH 7.2, mutant enzyme R310K	Methanosarcina thermophila
14.5	-	CoA	23°C, pH 7.2, mutant enzyme S309A	Methanosarcina thermophila
15.4	-	CoA	23°C, pH 7.2, mutant enzyme S309T	Methanosarcina thermophila
69	-	CoA	23°C, pH 7.2, mutant enzyme R310Q	Methanosarcina thermophila
230	-	CoA	23°C, pH 7.2, mutant enzyme R310A	Methanosarcina thermophila
2150	-	CoA	23°C, pH 7.2, mutant enzyme D316E	Methanosarcina thermophila
5190	-	CoA	23°C, pH 7.2, wild-type enzyme	Methanosarcina thermophila