Stereochemistry of methylene transfer involving 5,10-methylenetetrahydrofolate

Abstract

The stereochemistry of the transfer catalyzed by rabbit liver serine transhydroxymethylase (EC 2.1.2.1) of the prochiral hydroxymethyl group from serine to tetrahydrofolate to form 5,10-methylenetetrahydrofolate was studied. Initial kinetic studies on labeled 5,10-methylenetetrahydrofolate showed that nonenzymatic racemization of the prochiral methylene center was buffer dependent and was slow under the conditions employed. Specifically tritiated (3 R)- and (3 S)-serines were employed to study the transfer reaction. Reactions were carried out under various conditions and the stereochemistry of the methylene C of the 5,10-methylenetetrahydrofolate produced was determined. The enzyme was partially stereospecific for this transfer reaction, proceeding with a loss of about 50% of the stereochemical purity of the transferred C center. Possible mechanistic interpretations of this finding are discussed.