Identification of a Yeast Peroxisomal Member of the Family of AMP‐Binding Proteins

摘要: We established a reverse-genetic approach to identify peroxisomal proteins involved in fatty acid metabolism of Saccharomyces cerevisiae. Putative peripheral membrane were isolated by successive extraction purified peroxisomes and HPLC SDS/PAGE. Six identified peptide sequence analysis, including acyl-CoA oxidase trifunctional enzyme the β-oxidation system as well malate dehydrogenase 3 carnitine acetyltransferase. In addition two previously unknown putative identified, an 40-kDa protein which we named Pcs60p, both major constituent fraction. Pcs60p is encoded ORF Z36091 chromosome II from S. cerevisiae consists 543 amino acids with molecular mass 60.5 kDa. Biochemical, immunofluorescence microscopy immunocytochemical data confirmed that but also localized matrix. Consistent intraperoxisomal localization, consensus for peroxisomal-targeting signal 1 (PTS1) present at extreme C-terminus Pcs60p. Deletion studies revealed localization depends on presence this sequence. Expression highly inducible oleic acid, however, dispensable growth single carbon source. belongs family act via ATP-dependent covalent binding AMP their substrates shows highest degree similarity Escherichia coli long chain synthetase.