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http://dx.doi.org/10.5352/JLS.2008.18.8.1036

Kinetic and Chemical Mechanism of Pig Liver Methenyltetrahydrofolate Synthetase  

Cho, Yong-Kweon (Department of Biochemistry and Health Science, College of Natural Sciences, Changwon National University)
Publication Information
Journal of Life Science / v.18, no.8, 2008 , pp. 1036-1041 More about this Journal
Abstract
Methenyltetrahydrofolate synthetase extract was obtained from mouse liver and purified via $30{\sim}70%$ ammonium sulfate fractionation, Fast Q anion exchange and phenyl agarose chromatography. HPLC gel chromatography and SDS-polyacrylamide electrophoresis experiments showed that the enzyme is a monomer with molecular weight of 23 kDa. Optimum temperature and pH were $35^{\circ}C$ and 6.5, respectively. The enzyme was chemically modified only by tetranitromethane and 1-ethyl-3- (3-dimethyl aminopropyl)-carbodiimide (EDC), indicating that tyrosine and carboxylate are in the active site. pH studies showed that 2 tyrosines are involved in the binding of the substrates and a carboxylate in catalysis. Therefore, the chemical mechanism of the enzyme is likely that 2 tyrosines bind to ATP and 5-formylTHFand a carboxylate acts as a general base.
Keywords
Methenyltetrahydrofolate synthetase; folate metabolism; chemical mechanism;
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