title: Inborn errors of cobalamin metabolism: effect of cobalamin supplementation in culture on methylmalonyl CoA mutase activity in normal and mutant human fibroblasts.
abstract: We have examined the effect of addition of hydroxocobalamin to growth medium on the activity of the adenosylcobalamin-requiring enzyme methylmalonyl CoA mutase in normal human fibroblasts and in mutant human fibroblasts derived from patients with inherited methylmalonicacidemia. The mutant cell lines were assigned to four distinct genetic complementation groups (cbl A, cbl B, cbl C, and cbl D), each deficient in some step in the synthesis of adenosylcobalamin from hydroxocobalamin. After control cells were grown in cobalamin-supplemented medium, mutase holoenzyme activitiy increased markedly in a time- and concentration-dependent fashion. Growth in cobalamin-supplemented medium had no effect on mutase activity in some mutant lines belonging to the cbl B group, while activity increased severalfold in other cbl B mutants and in all cbl A, cbl C, and cbl D mutants examined, although mutase activity was still less than 10% of control. Comparison of mutase holoenzyme activity and total propionate pathway activity suggests that enhancement of mutase activity in mutant cells after cobalamin supplementation to values 5--10% of control may be sufficient to overcome the inherited metabolic block and to restore total pathway activity to normal.
mesh_terms: D000592:Amino Acid Metabolism, Inborn Errors / Q000235:genetics*; D002460:Cell Line; D003470:Culture Media; D014644:Genetic Variation; D006801:Humans; D007535:Isomerases / Q000378:metabolism*; D008314:Malonates / Q000378:metabolism*; D008764:Methylmalonic Acid / Q000378:metabolism*; D008765:Methylmalonyl-CoA Mutase / Q000378:metabolism*; D009154:Mutation; D014805:Vitamin B 12 / Q000494:pharmacology*
chemical_list: D003470:Culture Media; D008314:Malonates; D008764:Methylmalonic Acid; D007535:Isomerases; D008765:Methylmalonyl-CoA Mutase; D014805:Vitamin B 12