title: Characterization of trypsin inhibitors from Tora-mame seeds, one of the Japanese cultivars of Phaseolus vulgaris.
abstract: Four isoinhibitors against bovine pancreatic trypsin were purified from Phaseolus vulgaris(cv. Tora-mame) seeds by extraction with water(pH 2.0), ammonium sulfate fractionation, gel chromatography on Sephacryl S-200, trypsin-Sepharose gel affinity chromatography, and chromatofocusing. They inhibit both trypsin and chymotrypsin strongly. Their molecular masses are 85 kDa, estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Their isoelectric points range 5.09 to 4.46. They are high in the content of aspartic acid, serine, proline, and half-cystine but low in valine, methionine, tyrosine, and phenylalanine. Tryptophan is absent from them completely. They are bound to both trypsin and chymotrypsin with equimolar ratio, and have separate and independent binding sites for both proteases. Chemical modification showed that the inhibitors are of lysine type.
authors.affiliation: Department of Agricultural Chemistry, Faculty of Bioresources, Mie University, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Furuichi
authors.forename: H
authors.initials: H
authors.lastname: Umekawa
authors.forename: T
authors.initials: T
authors.lastname: Takahashi
mesh_terms: D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D002918:Chymotrypsin / Q000037:antagonists & inhibitors / Q000378:metabolism; D007887:Fabaceae / Q000201:enzymology*; D007526:Isoelectric Point; D007564:Japan; D008239:Lysine; D008970:Molecular Weight; D010940:Plant Proteins / Q000302:isolation & purification* / Q000378:metabolism; D010946:Plants, Medicinal; D013045:Species Specificity; D014357:Trypsin / Q000378:metabolism; D014361:Trypsin Inhibitors / Q000145:classification / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D000596:Amino Acids; D010940:Plant Proteins; D014361:Trypsin Inhibitors; D002918:Chymotrypsin; D014357:Trypsin; D008239:Lysine