title: The specificity in vivo of two distinct methionine aminopeptidases in Saccharomyces cerevisiae.
abstract: In Saccharomyces cerevisiae, the essential function of amino-terminal methionine removal is provided cotranslationally by two methionine aminopeptidases (MetAP1 and MetAP2). To examine the individual processing efficiency of each MetAP in vivo, we measured the degree of N-terminal methionine cleavage from a series of mutated glutathione-S-transferase (GST) proteins isolated from yeast wild-type, a map1 deletion strain, a map2 deletion strain, and a map1 deletion strain overexpressing the MAP2 gene. We found that MetAP1 plays the major role in N-terminal methionine removal in yeast. Both MetAPs were less efficient when the second residue was Val, and MetAP2 was less efficient than MetAP1 when the second residue was Gly, Cys, or Thr. These findings indicate that MetAP1 and MetAP2 exhibit different cleavage efficiencies against the same substrates in vivo. Interestingly, although methionine is considered a stabilizing N-terminal residue, we found that retention of the initiator methionine on the Met-Ala-GST mutant protein drastically reduced its half-life in vivo.
authors.affiliation: Edward A. Doisy Department of Biochemistry and Molecular Biology, St. Louis University School of Medicine, 1402 S. Grand Boulevard, St. Louis, Missouri 63104, USA.
authors.forename: Shaoping
authors.initials: S
authors.lastname: Chen
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Vetro
authors.forename: Yie-Hwa
authors.initials: YH
authors.lastname: Chang
mesh_terms: D019943:Amino Acid Substitution; D000626:Aminopeptidases / Q000235:genetics / Q000378:metabolism*; D004795:Enzyme Stability; D005982:Glutathione Transferase / Q000378:metabolism; D008715:Methionine / Q000378:metabolism*; D063208:Methionyl Aminopeptidases; D009154:Mutation; D010455:Peptides / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D010455:Peptides; D008715:Methionine; D005982:Glutathione Transferase; D000626:Aminopeptidases; D063208:Methionyl Aminopeptidases