蛋白质二硫键异构酶相关蛋白A的表达及性质的研究  

Expression and Characterization of Protein Disulfide Isomerase-related Protein A

作　　者：周海平[1] 贾翠娟[1] 张渝英[1] 

机构地区：[1]中国科学院微生物研究所,北京100080

出　　处：《生物工程学报》2004年第4期595-600,共6页Chinese Journal of Biotechnology

基　　金：国家自然科学基金资助项目 (No .3 9970 0 3 )~~

摘　　要：克隆了AspergillusnigerT2 1中的蛋白质二硫键异构酶相关蛋白A(PRPA)基因 ,并将它插入pET2 3b表达载体。在E .coli中表达时 ,PRPA占菌体总蛋白的 34%。经过超声破细胞、硫酸铵分级沉淀和离子交换层析获得了纯度大于 90 %的重组蛋白。PRPA有二硫键异构酶活性。在PRPA存在下 ,变性和还原的溶菌酶复性率和复性速度降低 ,电泳结果表明溶菌酶聚集增多。Protein disulfide isomerase related protein A (PRPA) was highly expressed (about 34%) in Escherichia coli by inserting the whole PRPA cDNA into the vector pET23b. After expression, the purified protein was acquired through ammonium fractional precipitation and Bio Rex 70 chromatography. PRPA shows low disulfide isomerase activity (only about 1/250 of that of hPDI), decreases the reactivation yield of denatured and reduced lysozyme either in redox and non redox Hepes buffer or redox PBS buffer and facilitates the aggregation of denatured and reduced lysozyme. Fluorescence spectra of PRPA indicate that PRPA has more hydrophobic groups at surface than that of hPDI, and which can be used to explain why PRPA has anti chaperone acti vity during the refolding of denatured and reduced lysozyme.

关 键 词：蛋白质二硫键异构酶相关蛋白A 抗分子伴侣 蛋白质重折叠 

分 类 号：Q55[生物学—生物化学]