Purification and properties of homoserine transacetylase from Bacillus polymyxa.
Homoserine transacetylase (EC 2.3.1.31), the first enzyme of methionine biosynthesis, has been purified to near homogeneity from extracts of a methionine auxotroph of Bacillus polymyxa. The enzyme is subject to rapid irreversible inactivation. Its half-life at 0 degrees is 15 min and much less at higher temperatures, but ethylene glycol affords some protection. In addition, Zn2+ reversibly inhibits the enzyme with a K-I of 3 muM. The enzyme has a molecular weight of about 40,000 and consists of a single polypeptide chain. Besides catalyzing the acetyl transfer from acetyl-CoA to L-homoserine, homoserine transacetylase promotes a homoserine-O-acetylhomoserine exchange reaction in the absence of CoA, suggesting the formation of an acetyl-enzyme intermediate. The results of kinetic studies are consistent with a ping-pong mechanism. Homoserine transacetylase is subject to multivalent feedback inhibition by L-methionine and S-adenosylmethionine. Analysis of the inhibition data and specificity studies suggest that the inhibitors bind to separate sites on the enzyme which are distinct from the active site. Inhibition is competitive with respect to both substrates, and the saturation curves for the inhibitors, as well as substrate saturation curves in the absence or presence of the inhibitors, are hyperbolic. The absence of cooperativity is, in fact, a property which would be expected in a monomeric allosteric enzyme such as homoserine transacetylase.
D000105:Acetyl Coenzyme A / Q000494:pharmacology; D000123:Acetyltransferases / Q000302:isolation & purification* / Q000378:metabolism; D000494:Allosteric Regulation; D001407:Bacillus / Q000201:enzymology*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D006207:Half-Life; D006714:Homoserine; D007700:Kinetics; D008715:Methionine / Q000494:pharmacology; D008970:Molecular Weight; D012436:S-Adenosylmethionine / Q000494:pharmacology; D013696:Temperature; D013997:Time Factors; D015032:Zinc / Q000494:pharmacology
D006714:Homoserine; D000105:Acetyl Coenzyme A; D012436:S-Adenosylmethionine; D008715:Methionine; D000123:Acetyltransferases; D015032:Zinc