Purification and some properties of cobalamin-dependent methionine synthase from rat liver

Cobalamin-dependent methionine synthase was purified from rat liver. The enzyme activity was separated into two peaks upon Mono-Q column chromatography. Peaks I and II of the enzyme, eluted in this order, were purified 18,000- and 44,000-fold in overall yields of 0.7 and 1.8%, respectively. Peak II methionine synthase, the major fraction, was homogeneous as judged by SDS-polyacrylamide gel electrophoresis. The enzyme was a large monomeric protein with an apparent molecular weight of 143,000 Da. Interconversion of the enzyme between the two peaks was not observed during purification procedures. The enzyme required S-adenosylmethionine and a reducing system for activity. Apparent K(m) values of the peak II enzyme for 5-methyltetrahydrofolate and homocysteine were 75 and 1.7 microM, respectively.