Ginsburg D, Zeheb R, Yang A Y, Rafferty U M, Andreasen P A, Nielsen L, Dano K, Lebo R V, Gelehrter T D
J Clin Invest. 1986 Dec;78(6):1673-80. doi: 10.1172/JCI112761.
Full-length cDNA for plasminogen activator inhibitor (PAI-1) was isolated from a human umbilical vein endothelial cell (HUVEC) lambda gt11 cDNA library. Three overlapping clones were identified by immunologic screening of 10(6) recombinant phage using a rabbit anti-human fibrosarcoma PAI-1 antiserum. The fusion proteins encoded by these three clones also react strongly with a monoclonal mouse anti-human fibrosarcoma PAI-1 antibody. By nucleotide sequence analysis, PAI-1 cDNA encodes a protein containing 402 amino acids with a predicted, nonglycosylated molecular mass of 45 kD. Identity of this material as authentic PAI-1 was confirmed by the presence of high level homology with the primary amino acid sequence of an internal peptide prepared from purified rat hepatoma PAI-1. The predicted amino acid sequence also reveals extensive homology with other members of the serine protease inhibitor gene family. Cultured HUVECs contain two PAI-1 mRNA species, both encoded by a single gene, differing by 1 kb in the 3' untranslated region. The PAI-1 gene is located on human chromosome 7.
从人脐静脉内皮细胞（HUVEC）λgt11 cDNA文库中分离出纤溶酶原激活物抑制剂（PAI-1）的全长cDNA。通过用兔抗人纤维肉瘤PAI-1抗血清对10⁶个重组噬菌体进行免疫筛选，鉴定出三个重叠克隆。这三个克隆编码的融合蛋白也能与小鼠抗人纤维肉瘤PAI-1单克隆抗体强烈反应。通过核苷酸序列分析，PAI-1 cDNA编码一种含有402个氨基酸的蛋白质，预测的非糖基化分子量为45 kD。通过与从纯化的大鼠肝癌PAI-1制备的内部肽段的一级氨基酸序列具有高度同源性，证实了该物质为真正的PAI-1。预测的氨基酸序列还显示与丝氨酸蛋白酶抑制剂基因家族的其他成员具有广泛的同源性。培养的HUVEC含有两种PAI-1 mRNA，均由单个基因编码，在3'非翻译区相差1 kb。PAI-1基因位于人类7号染色体上。
