利用大肠杆菌鞭毛展示的随机肽库筛选TNF-α拮抗肽  

Screening of TNF-α Antagonist Peptides from a Random Peptide Library Displayed with Escherichia coli Flagellar

作　　者：李朝[1] 周艳荣[1] 程度胜[1] 黄培堂[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071

出　　处：《生物工程学报》2002年第5期556-560,共5页Chinese Journal of Biotechnology

基　　金：军事医学科学院科研启动基金资助 (No .99110 8)~~

摘　　要：TNF α是一种在机体抗感染、抗肿瘤过程中发挥重要作用的细胞因子 ,其对机体具有保护和损伤两方面的作用 ,为了探讨新的抑制TNF α所致炎症损伤等反应的手段 ,构建了细菌鞭毛递呈的随机肽库 ,利用构建得到的肽库 ,进行TNF α特异性结合肽的筛选工作。经过 5轮筛选及DNA测序 ,共得到 6条小肽编码序列。其中 2条序列中含有一V N WG的相同序列框架。进行 6条序列与TNF α结合力的确证后 ,选择了其中的 4条肽序列进行人工化学合成、纯化及鉴定。利用L92 9细胞及MTT法对 4条小肽进行活性测定 ,检测其对TNF α的抑制活性。结果表明 ,在TNF α对L92 9细胞毒性为 30 %左右时 ,含同源序列框架的 2条肽可抑制 90 %左右的TNFTumor necrosis factor(TNF-α) plays an improtant role in the process of anti-infection and anti-cancer.It can both protect and make damage to the host.In order to find new way of inhibiting its host-damaging activity, An E.coli flagella displayed random peptide library was constructed and TNF-αantagonist peptides were screened using the peptide library. After 5 rounds of panning and DNA sequencing, six peptide sequences were obtained.Two of them(TBP2,TBP3) have the same sequence frame of V--N-WG.After primary comparation of TNF-α binding ability,four peptides were synthesised and purified with RP-HPLC.The activity of inhibiting TNF-α was detected with L929 cell and MTT method.The data show that TBP2 and TBP3 can inhibit 90% of TNF-α activity when TNF-αgives about 30% cell toxicity on L929.The two sequences have not been reported.

关 键 词：大肠杆菌鞭毛 随机肽库 筛选 TNF-α拮抗肽 

分 类 号：R346[医药卫生—基础医学]