热休克蛋白CpkB对nrhTNF体外复性的促进作用  

The promoting action of chaperonin CpkB on the renaturation of denatured nrhTNF in vitro

作　　者：颜真[1] 张英起[1] 王俊楼[1] 赵宁[1] 朱宝娥[1] 苏成芝[2] 

机构地区：[1]第四军医大学科研处生物技术中心,陕西西安710033 [2]第四军医大学科研处生物技术中心基础部生物化学与分子生物学教研室,陕西西安710033

出　　处：《第四军医大学学报》1999年第8期664-666,共3页Journal of the Fourth Military Medical University

基　　金：全军医药卫生科研基金!96Q088

摘　　要：目的：探讨嗜热菌热休克蛋白CpkB在体外与新型重组人肿瘤坏死因子（nrhTNF）蛋白质复性的关系．方法：在原核细胞中表达并纯化CpkB蛋白;将nrhTNF在8mol／I-W中变性，然后在不同浓度的CpkB存在下进行体外复性，以复性后nrhTNF活性检测指标进行分析结果：在一定浓度下CpkB对nrhTNF的复性有明显的促进作用，最高复性率达到75％，而不使用CpkB的nrhTNF复性率最高仅引％结论：CpkB具有分子伴侣作用，可促进变性蛋白恢复正常结构与功能。AIM: To study the possibility of using heat shockprotein CpkB from hyperthermophilic bacteria KODl to promote the renaturation of denatured recombinant new humantumor necrosis factor (nrhTNF) in vitro. METHODS: nrhTNF was denatured in & mol/l. Urea and then renatured in different concentrations of CpkB expressed and purifired fromE. colt JM 109 (pBVCpk ). The percentage of renaturationwas measured by the detection of nrhTNF activity in I-929cell line. RESULTS: CpkB, as a monomer, promoted the renaturatlon of nrhTNF when the ratio of CpkB to nrhTNF is10: I -- 25: l, the highest percentage of renaturation beingup to 75 in the experiment. While in negative control, thepercentage of renaturation is only 31. CONCLUSION: CpkBhas been proved to be a molecular chaperone, which can bindmalfolded protein and mediate the correct assembly ofpolypeptlde. CpkB can be applied in protein renaturation invitro.

关 键 词：热休克蛋白美 肿瘤坏死因子 复性 

分 类 号：Q789[生物学—分子生物学]