5-Methyltetrahydrofolate homocysteine cobalamin methyltransferase in human bone marrow and its relationship to pernicious anemia

Abstract

Dialyzed extracts from human bone marrow catalyze [5- 14C]methyltetrahydrofolate homocysteine transmethylation at slow but significant rates which can be detected by using substrate with a very high specific radioactivity. Enzymatic activity is associated with nucleated marrow cells rather than mature, nondividing erythrocytes. Extract transmethylase activities in 15 marrow specimens from patients without B-12 deficiency ranged from 157–1020 pmoles of [ Me- 14C]methionine formed/hr/10 7 nucleated cells. Catalysis is dependent on S-adenosyl- l-methionine and a flavin-reducing system, typical for the presence of a cobalamin (B-12) methyltransferase. No in vitro requirement for exogenous B-12 was observed except for the marrow extracts from two patients known to be B-12 deficient. One of these extracts was markedly stimulated by methyl-B-12 indicative that mostly apomethyltransferase was present. These tracer assays with cell-free extracts provide the first direct evidence that human bone marrow contains B-12 methyltransferase; they also afford further evidence for a 5-methyltetrahydrofolate trap in B-12 deficiency with its associated megaloblastic anemia. In addition, we have observed that in normal peripheral blood leukocytes the mononuclear fraction contains 10–30 times as much B-12 methyltransferase per nucleated cell as the polymorphonuclear granulocyte fraction.