title: Three novel subtilisin-trypsin inhibitors from Streptomyces: primary structures and inhibitory properties.
abstract: Three novel proteinaceous inhibitors, which had been identified as "Streptomyces subtilisin inhibitor-like (SIL) proteins" and exhibited trypsin inhibition in addition to strong inhibition toward subtilisin BPN', were purified from the culture broth of three Streptomyces strains: SIL10 from S. thermotolerans, SIL13 from S. galbus, and SIL14 from S. azureus. Their primary structures were determined by sequence analysis of intact SIL inhibitors and peptides obtained by enzymatic digestions of S-pyridylethylated SIL inhibitors. These inhibitors were composed of about 110 amino acids and existed as dimer proteins. The reactive site was identified as Lys-Gln for all three inhibitors by sequence analysis of their modified forms in which the reactive-site peptide bond was specifically cleaved by subtilisin BPN' under acidic conditions. Thus, their inhibition toward trypsin and subtilisin BPN' was due to the presence of a Lys residue at the P1 site. Inhibitor constants toward subtilisin BPN' and trypsin were also determined. These inhibitors showed relatively high sequence homology to other SSI-family inhibitors possessing a Lys residue at the P1 site, with amino acid replacements on their molecular surface.
authors.affiliation: Institute for Biomolecular Science, Gakushuin University, Tokyo.
authors.forename: M
authors.initials: M
authors.lastname: Terabe
authors.forename: S
authors.initials: S
authors.lastname: Kojima
authors.forename: S
authors.initials: S
authors.lastname: Taguchi
authors.forename: H
authors.initials: H
authors.lastname: Momose
authors.forename: K
authors.initials: K
authors.lastname: Miura
mesh_terms: D000595:Amino Acid Sequence; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D015842:Serine Proteinase Inhibitors / Q000302:isolation & purification*; D013302:Streptomyces / Q000378:metabolism*; D013381:Subtilisins / Q000037:antagonists & inhibitors*; D014361:Trypsin Inhibitors / Q000302:isolation & purification*
chemical_list: D015842:Serine Proteinase Inhibitors; D014361:Trypsin Inhibitors; D013381:Subtilisins