title: Pepsinogens and pepsins from Japanese seabass (Lateolabrax japonicus).
abstract: Three pepsinogens (PG1, PG2, PG3) were highly purified from the stomach of Japanese seabass (Lateolabrax japonicus) by ammonium sulfate fractionation, DEAE-Sephacel anion exchange column chromatography and Sephacryl S-200 gel-filtration. Two dimensional polyacrylamide gel electrophoresis (2D-PAGE) analysis revealed that the molecular masses of the three PGs were 35, 37, and 34kDa, and their isoelectric points were 5.3, 5.1, and 4.7, respectively. Zymography analysis showed that the three pepsinogens had different mobilities and enzymatic activities under native conditions. Pepsinogens converted into their active form pepsins under pH 2.0 by one-step pathway or stepwise pathway. All three pepsins were completely inhibited by pepstatin A, a typical aspartic proteinase inhibitor. The N-terminal amino acid sequences of the three pepsinogens were determined to the 30th, 30th and 28th amino acid residue and those of their corresponding active form pepsins were also determined to the 19th, 18th and 20th amino acid residue, respectively. All amino acid sequences of Japanese seabass PGs revealed high identities to reported fish and mammalian pepsinogens. The effective digestion of fish and shrimp muscular proteins by pepsins indicated their physiological function in the degradation of food proteins.
authors.affiliation: College of Biological Engineering, The Key Laboratory of Science and Technology for Aquaculture and Food Safety, Jimei University, Jimei, Xiamen, 361021, China.
authors.forename: Min-Jie
authors.initials: MJ
authors.lastname: Cao
authors.forename: Wei-Qin
authors.initials: WQ
authors.lastname: Chen
authors.forename: Cui-Hong
authors.initials: CH
authors.lastname: Du
authors.forename: Asami
authors.initials: A
authors.lastname: Yoshida
authors.forename: Wei-Guang
authors.initials: WG
authors.lastname: Lan
authors.forename: Guang-Ming
authors.initials: GM
authors.lastname: Liu
authors.forename: Wen-Jin
authors.initials: WJ
authors.lastname: Su
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D008969:Molecular Sequence Data; D010434:Pepsin A / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D010435:Pepsinogens / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D010473:Perciformes / Q000378:metabolism*; D016415:Sequence Alignment; D013270:Stomach / Q000201:enzymology
chemical_list: D010435:Pepsinogens; D010434:Pepsin A