title: Yeast methionine aminopeptidase I can utilize either Zn2+ or Co2+ as a cofactor: a case of mistaken identity?
abstract: Yeast methionine aminopeptidase I (MetAP I) is one of two enzymes in Saccharomyces cerevisiae that is responsible for cotranslational cleavage of initiator methionines. It has previously been classified as a Co2+ metalloprotease in all prokaryotic and eukaryotic forms studied. However, treatment of recombinant apo-MetAP I with 12.5 microM Zn2+ produces an enzyme that is as active as that reconstituted with 200 microM Co2+. In the presence of physiological concentrations of reduced glutathione (GSH), Co-MetAP I is inactive, while the activity of Zn-MetAP I is increased more than 1.7-fold over Zn-MetAP I assayed in the absence of GSH. Given that the in vivo concentration of Zn2+ is at least 1,000-fold higher than that of Co2+, and that Co2+ is insoluble in physiological concentrations of GSH, it is probable that yeast MetAP I is actually a Zn2+ metalloprotease. Furthermore, unless there are extraordinary conditions that insulate or sequester them from this reducing milieu, that have yet to be identified, there are not likely to be any cytoplasmic enzymes that use free Co2+.
authors.affiliation: Department of Physiology and Biophysics, College of Medicine, University of California, Irvine 92697, USA.
authors.forename: K W
authors.initials: KW
authors.lastname: Walker
authors.forename: R A
authors.initials: RA
authors.lastname: Bradshaw
mesh_terms: D000626:Aminopeptidases / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D001667:Binding, Competitive; D002412:Cations; D003035:Cobalt / Q000378:metabolism*; D004789:Enzyme Activation; D005656:Fungal Proteins / Q000187:drug effects / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism / Q000494:pharmacology; D063208:Methionyl Aminopeptidases; D011994:Recombinant Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D015003:Yeasts / Q000201:enzymology*; D015032:Zinc / Q000378:metabolism*
chemical_list: D002412:Cations; D005656:Fungal Proteins; D011994:Recombinant Proteins; D003035:Cobalt; D000626:Aminopeptidases; D063208:Methionyl Aminopeptidases; D005978:Glutathione; D015032:Zinc