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.
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
