title: Cloning and nucleotide sequence of opdA, the gene encoding oligopeptidase A in Salmonella typhimurium.
abstract: The opdA gene (formerly called optA) of Salmonella typhimurium encodes a metallopeptidase, oligopeptidase A (OpdA), first recognized by its ability to cleave and allow utilization of N-acetyl-L-Ala4 (E. R. Vimr, L. Green, and C. G. Miller, J. Bacteriol. 153:1259-1265, 1983). Derivatives of pBR328 carrying the opdA gene were isolated and shown to express oligopeptidase activity at levels approximately 100-fold higher than that of the wild type. These plasmids complemented all of the phenotypes associated with opdA mutations (failure to use N-acetyl-L-Ala4, defective phage P22 development, and diminished endopeptidase activity). The opdA region of one of these plasmids (pCM127) was defined by insertions of Tn1000 (gamma delta), and these insertions were used as priming sites to determine the nucleotide sequence of a 2,843-bp segment of the insert DNA. This region contained an open reading frame coding for a 680-amino-acid protein, the N terminus of which agreed with that determined for purified OpdA. This open reading frame contained both a sequence motif typical of Zn2+ metalloproteases and a putative sigma 32 promoter. However, no induction was detected upon temperature shift by using a beta-galactosidase operon fusion. The predicted OpdA sequence showed similarity to dipeptidyl carboxypeptidase, the product of the S. typhimurium gene dcp, and to rat metallopeptidase EC 3.4.24.15., which is involved in peptide hormone processing.
authors.affiliation: Department of Molecular Biology and Microbiology, School of Medicine, Case Western Reserve University, Cleveland, Ohio 44106.
authors.forename: C A
authors.initials: CA
authors.lastname: Conlin
authors.forename: C G
authors.initials: CG
authors.lastname: Miller
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D010450:Endopeptidases / Q000235:genetics*; D008666:Metalloendopeptidases / Q000235:genetics; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D012486:Salmonella typhimurium / Q000201:enzymology / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid; D015032:Zinc
chemical_list: D010450:Endopeptidases; C020071:dipeptidyl carboxypeptidase; D008666:Metalloendopeptidases; C057543:oligopeptidase A; D015032:Zinc