大肠杆菌T蛋白的聚合状态分析.pdfVIP

大肠杆菌T蛋白的聚合状态分析 王光柱，丁丁，刊、红颖，陈枢青‘(浙江大学药学院生物制药研究室，杭州310031) 摘要：目的解析大肠杆菌T蛋白的四级结构。方法利用分段克隆法克隆表达了T蛋白及其两个独立结构域分支酸变位酶 关键词：大肠杆菌T蛋白；分支酸变位酶；预苯酸脱氢酶；聚合状态 中图分类号：Q503 文献标识码：A 文章编号：1001—2494(2006)15—1190—04 of coli Status Escherichia Polymerization T·protein Analysis WANG Biopharmaceutics，Collegeof Guang-zhu，DINGDing，SUNHong—ying，CHENShu-qing“(脚硎删眦of 310031，China) Sciences，Zhej溉University，Hangzhou of Escherichiacoli．METHODS andPDHdo- Al塔Ⅱ认CT：oBlECll、，ETodescribethe structure from tertiary T-protein T-protein，CM maimwereclonedand SDSunderdenaturedconditionwere 11X M，determined 42×l∥，32×lo，and expressedseparately．‘11Ile by for tothetheoretical calculated 103 domainandCM wereidentical respeetively domain，which Mr．ne M，determinedby T-protein，PDH was 唧Cunderthenativeconditionwere63×103and25×103 forPDHdomainandCMdomain．Chemicalcross em— respectively linking todeterminethe statusof its in}聃_￡determination．REsIII腮％e doubled M，Wasover-range M，were ployed polymerizationT-protein，for forPI)HandCMdomainundernativecondition．A1lof andPDHare Wastestifiedboth}玎吃Candchemical T-protein，CM diin／2qer8，Which by ofCMtoCMandPDHtoPDHdo— isadimmeranditbonds adhesion cross-linkingexperiments．CONCLUSIONT-protein