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 the iron in heme-containing proteins. Cobalamin (vitamin B(12)) is structurally similar to heme and is a cofactor for methionine synthase, a key enzyme in folate metabolism. NO inhibits methionine synthase activity in vitro, but data concerning NO binding to cobalamin are controversial. We now show spectroscopically that NO reacts with all three valency states of cobalamin and that NO's inhibition of methionine synthase activity most likely involves its reaction with monovalent cobalamin. By following incorporation of the methyl moiety of [(14)C]methyltetrahydrofolic acid into protein, we show that NO inhibits methionine synthase activity in vivo, in cultured mammalian cells. The inhibition of methionine synthase activity disrupted carbon flow through the folate pathway as measured by decreased incorporation of [(14)C]formate into methionine, serine, and purine nucleotides. Homocysteine, but not cysteine, attenuated NO's inhibition of purine synthesis, providing further 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 NO 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 flow through the folate pathway.
Danishpajooh
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IO
Department of Medicine, University of California, San Diego, La Jolla, California 92093-0652, USA.
Gudi
T
T
Chen
Y
Y
Kharitonov
V G
VG
Sharma
V S
VS
Boss
G R
GR
D013766:5-Methyltetrahydrofolate-Homocysteine S-Methyltransferase / Q000037:antagonists & inhibitors*; D000818:Animals; D002244:Carbon / Q000378:metabolism*; D002460:Cell Line; D018920:Coculture Techniques; D006224:Cricetinae; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005492:Folic Acid / Q000378:metabolism*; D006801:Humans; D008715:Methionine / Q000096:biosynthesis; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000502:physiology*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D011685:Purine Nucleotides / Q000096:biosynthesis; D051381:Rats; D012694:Serine / Q000096:biosynthesis; D014805:Vitamin B 12 / Q000378:metabolism
D020030:Nitric Oxide Donors; D011685:Purine Nucleotides; D009569:Nitric Oxide; D012694:Serine; D002244:Carbon; D005492:Folic Acid; D008715:Methionine; D013766:5-Methyltetrahydrofolate-Homocysteine S-Methyltransferase; D014805:Vitamin B 12; D019331:NG-Nitroarginine Methyl Ester