A human vitamin B12 trafficking protein uses glutathione transferase activity for processing alkylcobalamins.
Pathways for tailoring and processing vitamins into active cofactor forms exist in mammals that are unable to synthesize these cofactors de novo. A prerequisite for intracellular tailoring of alkylcobalamins entering from the circulation is removal of the alkyl group to generate an intermediate that can subsequently be converted into the active cofactor forms. MMACHC, a cytosolic cobalamin trafficking chaperone, has been shown recently to catalyze a reductive decyanation reaction when it encounters cyanocobalamin. In this study, we demonstrate that this versatile protein catalyzes an entirely different chemical reaction with alkylcobalamins using the thiolate of glutathione for nucleophilic displacement to generate cob(I)alamin and the corresponding glutathione thioether. Biologically relevant thiols, e.g. cysteine and homocysteine, cannot substitute for glutathione. The catalytic turnover numbers for the dealkylation of methylcobalamin and 5'-deoxyadenosylcobalamin by MMACHC are 11.7 +/- 0.2 and 0.174 +/- 0.006 h(-1) at 20 degrees C, respectively. This glutathione transferase activity of MMACHC is reminiscent of the methyltransferase chemistry catalyzed by the vitamin B(12)-dependent methionine synthase and is impaired in the cblC group of inborn errors of cobalamin disorders.
Kim
Jihoe
J
Department of Biological Chemistry, University of Michigan Medical Center, Ann Arbor, Michigan 48109-5066, USA.
Hannibal
Luciana
L
Gherasim
Carmen
C
Jacobsen
Donald W
DW
Banerjee
Ruma
R
D000473:Alkanes / Q000378:metabolism; D001692:Biological Transport; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002384:Catalysis; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D003037:Cobalt Radioisotopes; D003038:Cobamides / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism*; D006801:Humans; D008954:Models, Biological; D009154:Mutation; D010088:Oxidoreductases; D021241:Spectrometry, Mass, Electrospray Ionization; D013438:Sulfhydryl Compounds / Q000378:metabolism; D014805:Vitamin B 12 / Q000031:analogs & derivatives* / Q000378:metabolism*
D000473:Alkanes; D002352:Carrier Proteins; D003037:Cobalt Radioisotopes; D003038:Cobamides; D013438:Sulfhydryl Compounds; C019476:mecobalamin; C508359:MMACHC protein, human; D010088:Oxidoreductases; D005982:Glutathione Transferase; C000913:cobamamide; D005978:Glutathione; D014805:Vitamin B 12