Characterization of a complementary deoxyribonucleic acid coding for human and bovine plasminogen.
Characterization of a complementary deoxyribonucleic acid coding for human and bovine plasminogen.
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编码人和牛纤溶酶原的互补脱氧核糖核酸的表征。
DOI：
10.1021/bi00313a035
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发表时间：
1984
期刊：
影响因子：
2.9
通讯作者：
Davie,EW
中科院分区：
文献类型：
--
作者：
Malinowski,DP;Sadler,JE;Davie,EW
Douglas P. Malinowski、J. Evan Sadler 和 Earl W. Davie* 摘要：在 pBR322 中利用牛肝 mRNA 构建了 cDNA 文库，该文库通过多核糖体免疫沉淀法富集了纤溶酶原 mRNA。然后从富集的牛 mRNA 中制备 A32p 标记的单链 cDNA，并用作筛选 cDNA 文库的探针。通过测试保护杂交的 32P 标记的 cDNA 免受 SI 核酸酶消化的克隆来进行筛选。发现的最长克隆长581个碱基对，编码牛纤溶酶原C端107个氨基酸、246个核苷酸的3'非编码区和多聚(A)尾。然后使用牛 cDNA 克隆作为探针来筛选包含 18 000 个重组体的人肝脏 cDNA 文库。发现六种分离物含有人纤溶酶原序列。由1851个碱基对组成的最长克隆是一种糖蛋白(MT～92000)，其参与纤维蛋白溶解的最后阶段(Robbins，1982)。在这些反应过程中，纤溶酶原转化为纤溶酶，这是一种水解纤维蛋白凝块的丝氨酸蛋白酶。纤溶酶原通过多种激活剂转化为纤溶酶，包括组织纤溶酶原激活剂、尿激酶或纤溶酶原和链激酶的复合物。在纤溶酶原转化为纤溶酶的过程中，单链前体分子中 Arg-560 和 Val-561 之间的特定内部肽键被裂解。纤溶酶由通过二硫键连接在一起的两条多肽链组成。该酶的丝氨酸蛋白酶部分位于蛋白质的羧基末端区域（Groskopf 等人，1969；Robbins 等人，1973）。该区域与参与血液凝固的丝氨酸蛋白酶以及参与其他生理过程（例如胰腺消化酶）的丝氨酸蛋白酶显示出相当大的氨基酸序列同源性（Davie等人，1979）。
Douglas P. Malinowski, J. Evan Sadler, and Earl W. Davie* abstract: A cDNA library was constructed in pBR322from bovine liver mRNA that was enriched for plasminogen mRNA by polysome immunoprecipitation. A32p-labeled singlestranded cDNA was then prepared from the enriched bovine mRNA and employed as a probe to screen the cDNA library. The screening was carried out by testing for clones that protect the hybridized 32P-labeled cDNA from SI nuclease digestion. The longest clone that was found was 581base pairs in length and coded for the C-terminal 107 amino acids of bovine plasminogen, a 3'noncoding region of 246 nucleotides and a poly (A) tail. Thebovine cDNA clone was then used as a probe to screen a human liver cDNA library of 18 000 recombinants. Six isolates were found to contain human plasminogen se-quences. The longest clone consisted of 1851 base pairsI^ asminogen is a glycoprotein (MT~ 92 000) that participates in the final stages of fibrinolysis (Robbins, 1982). During these reactions, plasminogen is converted to plasmin, a serine pro-tease that hydrolyzes fibrin clots. Plasminogen is converted to plasmin by a number of activators, including tissue plas-minogen activator, urokinase, or a complex of plasminogen and streptokinase. During the conversion of plasminogen to plasmin, a specific internal peptide bond between Arg-560 and Val-561 is cleaved in the single-chain precursor molecule. Plasmin is composed of two polypeptide chains held together by disulfide bonds. The serine protease portion of the enzyme is located in the carboxyl-terminal region of the protein (Groskopf et al., 1969; Robbins et al., 1973). This region shows considerableamino acid sequence homology with the serine proteases involved in blood coagulation, as well as those involved in other physiological processes such as the digestive enzymes of the pancreas (Davie et al., 1979).
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影响因子：
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