Kageyama T, Takahashi K
J Biochem. 1976 Mar;79(3):455-68. doi: 10.1093/oxfordjournals.jbchem.a131089.
Five pepsinogens were purified from gastric mucosa of Japanese monkey by DEAE-cellulose column chromatography and Sephadex G-100 gel filtration. Each was shown to be homogeneous by polyacrylamide disc gel electrophoresis. They were designated as pepsinogens I, II, III-1, III-2, and III-3, respectively, based on the elution profile on DEAE-cellulose chromatography. The molecular weights of pepsinogens I and II were 48,000 and 43,000, respectively, and those of the other three were 40,000. Each pepsinogen was converted to pepsin [EC 3.4.23.1] by acidification, and some characteristics, e.g. the pH dependence of activity, sensitivity to various inhibitors, stability to alkali, and hydrolytic activity toward N-acetyl-L-phenylalanyl-3,5-diiodo-L-tyrosine (APDT), were determined. The characteristics of pepsins I and II were the same, and those of pepsins III-1, III-2, and III-3 were similar. Pepsin III-3 showed high stability to alkali (pH 8.0), while the others were less stable. Each pepsin hydrolyzed APDT and was inhibited by acid protease-specific inhibitors, e.g. pepstain, diazoacetyl-DL-norleucine methyl ester (DAN), 1,2-epoxy-3-(p-nitrophenoxy)propane (EPNP), and p-bromophenacyl bromide. The compositions of pepsins I and II were the same, indicating that they are the same protein, and those of pepsins III-1, III-2, and III-3 resembled that of human pepsin. The diversity of pepsinogens and pepsins is discussed in comparison with pepsinogens and pepsins from other animals.
通过DEAE - 纤维素柱色谱和Sephadex G - 100凝胶过滤从日本猕猴胃黏膜中纯化出5种胃蛋白酶原。经聚丙烯酰胺圆盘凝胶电泳显示,每种胃蛋白酶原均为纯品。根据它们在DEAE - 纤维素色谱上的洗脱图谱,分别将其命名为胃蛋白酶原I、II、III - 1、III - 2和III - 3。胃蛋白酶原I和II的分子量分别为48,000和43,000,其他三种的分子量为40,000。每种胃蛋白酶原经酸化后转化为胃蛋白酶[EC 3.4.23.1],并测定了一些特性,如活性的pH依赖性、对各种抑制剂的敏感性、对碱的稳定性以及对N - 乙酰 - L - 苯丙氨酰 - 3,5 - 二碘 - L - 酪氨酸(APDT)的水解活性。胃蛋白酶I和II的特性相同,胃蛋白酶III - 1、III - 2和III - 3的特性相似。胃蛋白酶III - 3对碱(pH 8.0)具有较高的稳定性,而其他胃蛋白酶稳定性较差。每种胃蛋白酶均可水解APDT,并被酸性蛋白酶特异性抑制剂如胃蛋白酶抑制剂、重氮乙酰 - DL - 正亮氨酸甲酯(DAN)、1,2 - 环氧 - 3 -(对硝基苯氧基)丙烷(EPNP)和对溴苯甲酰溴抑制。胃蛋白酶I和II的组成相同,表明它们是同一种蛋白质,胃蛋白酶III - 1、III - 2和III - 3的组成与人胃蛋白酶的组成相似。并与其他动物的胃蛋白酶原和胃蛋白酶相比较,讨论了胃蛋白酶原和胃蛋白酶的多样性。