title: The amino acid sequence of a 20 kDa bifunctional subtilisin/alpha-amylase inhibitor from bran [correction of brain] of rice (Oryza sativa L.) seeds.
abstract: A 20 kDa bifunctional inhibitor of the microbial proteinase, subtilisin, and the alpha-amylase from the larvae of the red flour beetle (Tribolium castaneum) was purified from bran of rice seeds by saline extraction, precipitation with ammonium sulphate, ion-exchange chromatography on DEAE-Cellulose and Toyopearl CM-650, and preparative HPLC on Vydac C18. The complete primary structure was determined by automatic degradation of the intact, reduced and S-alkylated protein, and by manual DABITC/PITC micro-sequencing of peptides obtained from the protein following separate enzymic digestions with trypsin, pepsin, chymotrypsin, elastase and the protease from S. aureus V8. The protein sequence, which contained 176 residues, showed strong homology with similar bifunctional inhibitors previously isolated from wheat and barley which are related to the Kunitz family of proteinase inhibitors from legume seeds.
authors.affiliation: Hokuriku National Agricultural Experimental Station, Ministry of Agriculture Forestry and Fisheries, Niigata, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Ohtsubo
authors.forename: M
authors.initials: M
authors.lastname: Richardson
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002852:Chromatography, Ion Exchange; D001517:Coleoptera / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D007814:Larva; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012275:Oryza / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry* / Q000302:isolation & purification; D012639:Seeds / Q000737:chemistry; D012689:Sequence Homology, Nucleic Acid; D013381:Subtilisins / Q000037:antagonists & inhibitors*; D000516:alpha-Amylases / Q000037:antagonists & inhibitors*
chemical_list: D010940:Plant Proteins; C076389:subtilisin inhibitor p20, Oryza sativa; D000516:alpha-Amylases; D013381:Subtilisins