Nitric oxide inhibits methionine synthase activity in vivo and disrupts carbon flow through the folate pathway

Many of nitric oxide's biological effects are mediated via NO binding to th e iron in heme-containing proteins. Cobalamin (vitamin B-12) is structurall y similar to heme and is a cofactor for methionine synthase, a key enzyme i n folate metabolism, NO inhibits methionine synthase activity in vitro, but data concerning NO binding to cobalamin are controversial. We now show spe ctroscopically that NO reacts with all three valency states of cobalamin an d that NO's inhibition of methionine synthase activity most likely involves its reaction with monovalent cobalamin, By following incorporation of the methyl moiety of [C-14]methyltetrahydrofolic acid into protein, we show tha t NO inhibits methionine synthase activity in vivo, in cultured mammalian c ells. The inhibition of methionine synthase activity disrupted carbon flow through the folate pathway as measured by decreased incorporation of [C-14] formate into methionine, serine, and purine nucleotides, Homocysteine, but not cysteine, attenuated NO's inhibition of purine synthesis, providing fur ther evidence that NO was acting through methionine synthase inhibition. NO 's effect was observed both when NO donors were added to cells and when NO was produced physiologically in co-culture experiments. Treating cells with an MO synthase inhibitor increased formate incorporation into methionine, serine, and purines and methyl-tetrahydrofolate incorporation into protein. Thus, physiological concentrations of NO appear to regulate carbon flaw th rough the folate pathway.