{"title":"组织型纤溶酶原激活物突变体的构建、表达与表征。","authors":"S Liu, P Huang, C Huang","doi":"","DOIUrl":null,"url":null,"abstract":"<p><p>Three tissue-type plasminogen activator (t-PA) mutants were constructed by recombinant and site-directed mutagenesis techniques. They are del(296-302) with deletion of PAI-1 binding site, N117Q/N184Q with deglycosylation of K1 and K2 domains, and their combination mutant designated as GGI. Then these three mutants were successfully transiently expressed in COS-7 cells, and GGI was further stably expressed in CHO cells. The biological characterization of the expression products indicated that del(296-302) and GGI possessed the resistance to inhibition by PAI-1. In addition, the specific activity of GGI was increased by about 46%, the plasma half-life was prolonged by about one fold, while its affinity for fibrin was not affected.</p>","PeriodicalId":21648,"journal":{"name":"Science in China. Series B, Chemistry, life sciences & earth sciences","volume":"38 11","pages":"1341-8"},"PeriodicalIF":0.0000,"publicationDate":"1995-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Construction, expression and characterization of tissue-type plasminogen activator mutants.\",\"authors\":\"S Liu, P Huang, C Huang\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Three tissue-type plasminogen activator (t-PA) mutants were constructed by recombinant and site-directed mutagenesis techniques. They are del(296-302) with deletion of PAI-1 binding site, N117Q/N184Q with deglycosylation of K1 and K2 domains, and their combination mutant designated as GGI. Then these three mutants were successfully transiently expressed in COS-7 cells, and GGI was further stably expressed in CHO cells. The biological characterization of the expression products indicated that del(296-302) and GGI possessed the resistance to inhibition by PAI-1. In addition, the specific activity of GGI was increased by about 46%, the plasma half-life was prolonged by about one fold, while its affinity for fibrin was not affected.</p>\",\"PeriodicalId\":21648,\"journal\":{\"name\":\"Science in China. Series B, Chemistry, life sciences & earth sciences\",\"volume\":\"38 11\",\"pages\":\"1341-8\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1995-11-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Science in China. Series B, Chemistry, life sciences & earth sciences\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Science in China. Series B, Chemistry, life sciences & earth sciences","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Construction, expression and characterization of tissue-type plasminogen activator mutants.
Three tissue-type plasminogen activator (t-PA) mutants were constructed by recombinant and site-directed mutagenesis techniques. They are del(296-302) with deletion of PAI-1 binding site, N117Q/N184Q with deglycosylation of K1 and K2 domains, and their combination mutant designated as GGI. Then these three mutants were successfully transiently expressed in COS-7 cells, and GGI was further stably expressed in CHO cells. The biological characterization of the expression products indicated that del(296-302) and GGI possessed the resistance to inhibition by PAI-1. In addition, the specific activity of GGI was increased by about 46%, the plasma half-life was prolonged by about one fold, while its affinity for fibrin was not affected.
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