Primary structure of two distinct rat pancreatic preproelastases determined by sequence analysis of the complete cloned messenger ribonucleic acid sequences.
Primary structure of two distinct rat pancreatic preproelastases determined by sequence analysis of the complete cloned messenger ribonucleic acid sequences.
复制标题
通过完整克隆的信使核糖核酸序列的序列分析确定两种不同的大鼠胰腺前原弹性蛋白酶的一级结构。
DOI：
10.1021/bi00535a053
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发表时间：
1982
期刊：
影响因子：
2.9
通讯作者：
Rutter,WJ
中科院分区：
文献类型：
--
作者：
MacDonald,RJ;Swift,GH;Quinto,C;Swain,W;Pictet,RL;Nikovits,W;Rutter,WJ
雷蒙德·麦克唐纳，1”高尔文·H。Swift，1 Carmen Quinto，** William Swain，Raymond L. Pictet，x William Nikovits和William J. Rutter摘要：通过重组DNA技术克隆了两种大鼠胰腺弹性蛋白溶解酶的mRNA序列，并测定了其核苷酸序列。大鼠弹性蛋白酶I mRNA长1113个核苷酸，加上一个poly（A）尾，编码266个氨基酸的前弹性蛋白酶原。预测的大鼠弹性蛋白酶I的活性形式的氨基酸序列与猪弹性蛋白酶1具有84%的同源性。决定猪弹性蛋白酶1底物特异性的关键氨基酸残基保留在大鼠酶中。酶原的激活肽似乎与其他哺乳动物胰腺丝氨酸蛋白酶的激活肽无关。弹性蛋白酶I的mRNA定位于腺泡细胞的粗面内质网中，正如外分泌分泌酶合成位点所预期的那样。大鼠弹性蛋白酶Ⅱ mRNA全长910个核苷酸，加上一个poly（A）尾，编码271个氨基酸的前酶原。氨基酸序列与猪弹性蛋白酶1（58%序列同一性）比与其它胰腺丝氨酸蛋白酶（33-39%序列同一性）更密切相关。基于定义底物结合裂隙的关键氨基酸残基的底物偏好预测与哺乳动物胰腺弹性蛋白酶2观察到的广泛特异性一致。活化肽类似于胰凝乳蛋白酶原的活化肽，并且保留可用于与内部保守半胱氨酸残基形成二硫键的N-末端半胱氨酸。哺乳动物的外分泌胰腺合成、储存和分泌大约15种酶和酶原用于肠内消化。这些分泌蛋白占腺体蛋白质合成的80%以上（Jamieson & Palade，1967; VanNest等人，1980年）。胰腺分泌物的主要部分是至少七种丝氨酸蛋白酶的家族：胰凝乳蛋白酶A和B、两种胰蛋白酶、弹性蛋白酶1和2、1和激肽释放酶。该基因家族的成员可能通过从共同的祖先原基因进化而相关（Neurath等人，1967; deHaen等人，1975年），并保留了类似的结构，大小和功能。作为分泌蛋白，丝氨酸蛋白酶预期最初作为前体蛋白合成（Devillers-Thiery等人，1975; Rutter等人，1978）与氨基末端信号肽（Blobel & Sabatini，1971），其规定了聚f的结合。（RJM和GHS），德克萨斯州达拉斯，达拉斯，德克萨斯州75235的德克萨斯大学健康科学中心，以及生物化学和生物物理学系（RJM、CQ、WS、RLP、WN和WJR），加州大学，弗朗西斯科，加州94143。1981年9月9日收到。这项工作得到了国家科学基金会（PCM 8006231，RJM）和国家卫生研究所（AM 21344，WJR）的支持。弗吉尼亚·拉森比·奥哈拉研究员。现地址：墨西哥莫雷洛斯，库埃尔纳瓦卡，Centro de Figacion de Nitrogeno x现地址：Institute de Biologie Moleculaire，Universite巴黎VII，Tour 43，75221巴黎Cedex 05，法国。
Raymond J. MacDonald, 1" Galvin H. Swift, 1 Carmen Quinto,** William Swain, Raymond L. Pictet, x William Nikovits, and William J. Rutter abstract: The mRNA sequences for two rat pancreatic elastolytic enzymes have been cloned by recombinant DNA technology and their nucleotide sequences determined. Rat elastase I mRNA is 1113 nucleotides in length, plus a poly (A) tail, and encodes a preproelastase of 266 amino acids. The amino acid sequence of the predicted active form of rat elastase I is 84% homologous to porcine elastase 1. Key amino acid residues involvedin determining substrate specificity of porcine elastase 1 are retained in the rat enzyme. The activation peptide of the zymogen does not appear related to that of other mammalian pancreatic serine proteases. The mRNA for elastase I is localized in the rough endoplasmic reticulum of acinar cells, as expected for the site of synthesis of an exocrine secretory enzyme. Rat elastase II mRNA is 910nucleotides in length, plus a poly (A) tail, and encodes a preproenzyme of 271 amino acids. The amino acid sequence is more closely related to porcine elastase 1 (58% sequence identity) than to the other pancreatic serine proteases (33-39% sequence identity). Predictions of substrate preference based upon key amino acid residues that define the substrate binding cleft are consistent with the broad specificity observed for mammalian pancreatic elastase2. The activation peptide is similar to that of the chymotrypsinogens and retains an N-terminal cysteine available to form a disulfide link to an internal conserved cysteine residue. e mammalian exocrine pancreas synthesizes, stores, and secretes approximately 15 enzymes and proenzymes for in-testinal digestion. These secretory proteins account for greater than 80% of the protein synthesis of the gland (Jamieson & Palade, 1967; VanNest et al., 1980). A dominant fraction of the pancreatic secretions is a family of at least seven serine proteases: chymotrypsins A and B, two trypsins, elastases 1 and 2, 1 and kallikrein. The members of this gene family are probably related by evolution from a common ancestral pro-tease (Neurath et al., 1967; deHaen et al., 1975) and have retained similar structure, size, and function. As secretory proteins, the serine proteases are expected to be synthesized initially as precursor proteins (Devillers-Thiery et al., 1975; Rutter et al., 1978) with an amino-terminal signal peptide (Blobel & Sabatini, 1971) that specifies the binding of poly-f From the Division of MolecularBiology of the Department of Biochemistry (RJM and GHS), the University of Texas Health Science Center at Dallas, Dallas, Texas 75235, and the Department of Biochemistry and Biophysics (RJM, CQ, WS, RLP, WN, and WJR), The University of California, San Francisco, California 94143. Received September 9, 1981. This work was supported by the National Science Foundation (PCM 8006231 to RJM) and the National Insti-tutes of Health (AM21344 to WJR).* Virginia Lazenby O’Hara fellow.* Present address: Centro de Figacion de Nitrogeno, Cuernavaca, Morelos, Mexico. x Present address: Institute de Biologie Moleculaire, Universite Paris VII, Tour 43, 75221 Paris Cedex 05, France.