title: Proteinase yscD (oligopeptidase yscD). Structure, function and relationship of the yeast enzyme with mammalian thimet oligopeptidase (metalloendopeptidase, EP 24.15).
abstract: The yeast PRD1 gene, encoding proteinase yscD, was cloned by complementation of the prd1-6 point mutation. Sequencing of the gene revealed an open reading frame of 2.136 kb, encoding a protein of 712 amino acids with a calculated molecular mass of 81.8 kDa. The sequence HEGLG beginning at residue 501 represents the HEXXH motif, unique for the zinc metallo-peptidases. Sequence comparison revealed complete identity of the proteinase yscD gene with a recently published open reading frame of yeast chromosome III. We found 34.8% identity between proteinase yscD and rat metalloendopeptidase (thimet oligopeptidase, EP 24.15). Proteinase yscD hydrolyzes several chromogenic and fluorogenic peptides that are substrates of thimet oligopeptidase. N-[1-(RS)-carboxy-3-phenylpropyl]-Ala-Ala-Phe-p-aminobenzoic acid, a compound designed as specific inhibitor of EP 24.15, is also a strong inhibitor of the yeast enzyme. Proteinase yscD is a nonvacuolar enzyme. 3-5% of the total enzyme activity can be detected in the intermembrane space of mitochondria. In a mutant carrying a deletion of the PRD1 gene no proteinase yscD activity is detectable in the cytoplasm and in mitochondria of these cells. They do not show any grossly altered phenotype but exhibit a decrease in the intracellular degradation of peptides. This suggests a function of proteinase yscD in the late stages of protein degradation.
authors.affiliation: Institut für Biochemie, Universität Stuttgart, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Büchler
authors.forename: U
authors.initials: U
authors.lastname: Tisljar
authors.forename: D H
authors.initials: DH
authors.lastname: Wolf
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001407:Bacillus / Q000201:enzymology / Q000235:genetics; D001483:Base Sequence; D015825:Chromosomes, Fungal; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli; D017353:Gene Deletion; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D005838:Genotype; D008099:Liver / Q000201:enzymology; D008322:Mammals; D008666:Metalloendopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D017354:Point Mutation; D051381:Rats; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D003546:Cysteine Endopeptidases; C045096:proteinase yscD; D008666:Metalloendopeptidases; C022510:thimet oligopeptidase