Isolation of a human cDNA for alpha 2-thiol proteinase inhibitor and its identity with low molecular weight kininogen.

Isolation of a human cDNA for alpha 2-thiol proteinase inhibitor and its identity with low molecular weight kininogen.
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α2-硫醇蛋白酶抑制剂的人 cDNA 的分离及其与低分子量激肽原的同一性。

DOI:
10.1021/bi00319a005
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发表时间:
1984
期刊:
影响因子:
2.9
通讯作者:
Sasaki,M
Sasaki,M
中科院分区:
生物学3区
文献类型:
--
作者:
Ohkubo,I;Kurachi,K;Takasawa,T;Shiokawa,H;Sasaki,M

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Iwao Ohkubo,Kotoku Kurachi,* Toshihide Takasawa,Hiroyuki Shiokawa,and Makoto Sasaki摘要:含有从人肝脏mRNA制备的DNA插入片段的Xgtll cDNA文库已经用从新鲜血浆中分离的人α 2-巯基蛋白酶抑制剂的抗体筛选。从100万个噬菌体中分离出18个阳性克隆,并对每个克隆进行噬斑纯化。对其中一个噬菌体的cDNA插入物进行测序,并显示编码α 2-巯基蛋白酶抑制剂,如通过α 2-巯基蛋白酶抑制剂轻链的部分氨基酸序列所鉴定的。该cDNA插入片段含有1529个碱基对,编码完整的α 2-巯基蛋白酶抑制剂。它包括45个碱基对的5 '端非编码序列、1281个碱基对的前2-巯基蛋白酶抑制剂、一个终止密码子、160个碱基对的3'端非编码序列和40个碱基对的poly(A)尾。3 '端的非编码序列含有一个潜在的前体信使RNA加工和多聚腺苷酸化的识别位点(AATAAA),由该cDNA推导的α 2-巯基蛋白酶抑制剂的氨基酸序列与牛低分子量(LMW)激肽原的氨基酸序列具有惊人的相似性(总同源性为74%),包括两个内部重复序列和一个缓激肽的九肽序列。这些数据清楚地表明α 2-巯基蛋白酶抑制剂和LMW激酶原是相同的。这进一步得到了α 2-巯基蛋白酶抑制剂和LMW激肽原之间的免疫交叉反应性的支持。当将α 2-巯基蛋白酶抑制剂的氨基酸序列与几种低分子量巯基蛋白酶抑制剂的氨基酸序列进行比较时,
Iwao Ohkubo, Kotoku Kurachi,* Toshihide Takasawa, Hiroyuki Shiokawa, and Makoto Sasaki abstract: A Xgtll cDNA library containing DNA inserts prepared from human liver mRNA has been screened with an antibody to human «2-thiol proteinase inhibitor that was isolated from fresh plasma. Eighteen positive clones were isolated from one million phage, and each was plaque purified. The cDNA insert of one of these phage was sequenced and shown to code for «2-thiolproteinase inhibitor as identified by a partial amino acid sequence of the light chain of a2-thiol proteinase inhibitor. This cDNA insertcontained 1529 base pairs coding for thecomplete a2-thiol proteinase inhibitor. It included 45 base pairs of 5'noncoding sequence, 1281 base pairs that code for pre «2-thiol proteinase inhibitor, a stop codon, 160 base pairs of 3'noncoding sequence, and 40 base pairs of poly (A) tail. The noncoding sequence on the 3'end contained a potential recognition site (AATAAA) for processing and polyadenylation of precursor messenger RNA.The amino acidsequence of a2-thiol proteinase inhibitor de-duced from the cDNA showed a striking similarity (overall homology at 74%) to that of bovine low molecular weight (LMW) kininogen, including two internally repeated sequences and a nonapeptide sequence of bradykinin. These data clearly indicated that a2-thiol proteinase inhibitor and LMW kini-nogen are identical. This was further supported by immu-nological cross-reactivity between «2-thiol proteinase inhibitor and LMW kininogen. When the amino acidsequence of «2-thiol proteinase inhibitor was compared with those for several lowmolecular weight thiol proteinase inhibitors, in-
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