title: Isolation and characterization of pepsinogen from Trimeresurus flavoviridis (Habu snake).
abstract: Pepsinogen was isolated from the gastric mucosa of Trimeresurus flavoviridis (Habu snake) by DEAE-cellulose and DEAE-Sepharose ion-exchange chromatographies, and Sephacryl S-200 gel-chromatography. The yield calculated from the crude extract was 29% with 6.2-fold purification. The purified pepsinogen gave a single band on both native- and SDS-PAGE. As no other active enzyme was detected on the chromatographies, it was concluded that the Habu snake has one major pepsinogen. The molecular mass of the pepsinogen was estimated to be 38 kDa by SDS-PAGE. The sequence of the N-terminal 26 amino acid residues was determined and compared with those of other pepsinogens. The N-terminal structure of Habu snake pepsinogen was more homologous with those of mammalian pepsinogens C than those of mammalian pepsinogens A. The pepsinogen was rapidly converted to pepsin by way of an intermediate form induced by acidification. The optimum pH of Habu snake pepsin for bovine hemoglobin was 1.5-2.0, and it retained full activity at pH 6.2 and 30 degrees C on incubation for 30 min. The optimum temperature for the snake pepsin was 50 degrees C and it was stable at 40 degrees C on incubation for 10 min. The proteolytic activity of the pepsin toward bovine hemoglobin was about two times higher than that of porcine pepsin A, however, the activity toward oxidized bovine insulin B-chain was lower than that of porcine pepsin A, and it did not hydrolyze oligopeptides. The specificity for oxidized bovine insulin B-chain of the pepsin was different from that of porcine pepsin A. Habu snake pepsin was inhibited by pepstatin A but not by serine, cysteine, or metallo protease inhibitors.
authors.affiliation: Department of Chemistry, Faculty of Science, Kagoshima University, Kagoshima 890-0065, Japan. Yonezawa@sci.kagoshima-u.ac.jp
authors.forename: H
authors.initials: H
authors.lastname: Yonezawa
authors.forename: T
authors.initials: T
authors.lastname: Nonaka
authors.forename: T
authors.initials: T
authors.lastname: Uchikoba
authors.forename: S
authors.initials: S
authors.lastname: Hattori
authors.forename: M
authors.initials: M
authors.lastname: Ohno
authors.forename: M
authors.initials: M
authors.lastname: Kaneda
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D005753:Gastric Mucosa / Q000201:enzymology*; D008969:Molecular Sequence Data; D010434:Pepsin A / Q000037:antagonists & inhibitors / Q000378:metabolism*; D020116:Pepsinogen A / Q000737:chemistry*; D011480:Protease Inhibitors / Q000494:pharmacology; D011499:Protein Processing, Post-Translational; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D017841:Trimeresurus
chemical_list: D011480:Protease Inhibitors; D020116:Pepsinogen A; D010434:Pepsin A