New subtilisin-trypsin inhibitors produced by Streptomyces: primary structures and their relationship to other proteinase inhibitors from Streptomyces.
Three new proteinaceous inhibitors of trypsin and subtilisin of the Streptomyces subtilisin inhibitor (SSI)-like (SIL) protein family were isolated and purified from culture media of Streptomyces strains; SIL5 from S. fradiae, SIL7 from S. ambofaciens and SIL12 from S. hygroscopicus. Their complete amino-acid sequences were determined by sequence analysis of the intact SIL proteins and peptides obtained by enzymatic digestion of S-pyridylethylated proteins. SIL7 showed high sequence similarity to other Arg-possessing SSI-family inhibitors at the P1 site. SIL12 is unique in having a two-residue insertion in the flexible loop region. Based on the amino-acid sequences of these inhibitors and other SSI-family inhibitors whose sequences have already been determined, the phylogenetic relationship of SSI-family inhibitors and Streptomyces strains was considered. Among about 110 amino-acid residues possessed by SSI-family inhibitors, 28 are completely conserved. The contribution of these conserved residues to the function and stability of the inhibitor molecules is discussed on the basis of the results obtained from mutational analysis of SSI and its crystal structure.
Terabe
M
M
Institute for Biomolecular Science, Gakushuin University, Tokyo, Japan.
Kojima
S
S
Taguchi
S
S
Momose
H
H
Miura
K
K
D000595:Amino Acid Sequence; D002848:Chromatography, DEAE-Cellulose; D017124:Conserved Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D008961:Models, Structural; D008969:Molecular Sequence Data; D010802:Phylogeny; D011480:Protease Inhibitors / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D017433:Protein Structure, Secondary; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism*; D013381:Subtilisins / Q000037:antagonists & inhibitors*; D014361:Trypsin Inhibitors / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification
D011480:Protease Inhibitors; D014361:Trypsin Inhibitors; D013381:Subtilisins