从噬菌体随机七肽库中筛选hERG钾通道亲和肽的研究  

Screening of peptides binding to hERG potassium channel from 7-mer random phage displayed peptide library

作　　者：孙洪良[1,2] 魏晓莉[1] 于金梅[1] 华南[1] 刘晓燕[1] 郑建全[1] 

机构地区：[1]军事医学科学院毒物药物研究所,北京100850 [2]海军机关门诊部,北京100027

出　　处：《军事医学》2011年第7期513-516,558,共5页Military Medical Sciences

摘　　要：目的获得与hERG钾通道特异性结合的功能性亲和肽。方法以hERG1a亚基第3个细胞外环(L3)为靶蛋白分子,对噬菌体随机七肽库进行亲和筛选,ELISA法检测噬菌体克隆与靶蛋白的亲和性,使用全自动膜片钳技术观察亲和肽的活性。结果与结论经过3轮亲和筛选后,随机挑选15个噬菌体克隆对其亲和性做ELISA鉴定,均显示较强的阳性结果。将上述阳性克隆进行DNA测序得到3种不同小肽序列。电生理学研究表明,合成的3个小肽均能显著抑制hERG钾电流,为靶向hERG钾通道抗肿瘤研究提供了新的研究思路。Objective To obtain the peptides with binding specifically to hERG potassium channel from 7-mer random phage display peptide library.Methods The third extra cellular loop of hERG1a（L3） was used as the target to screen its binding peptides from 7-mer random phage display peptide library,and positive clones were isolated,followed by sequence analysis and activity evaluation.Results and Conclusion After three rounds of biopanning,three phage clones were obtained with different DNA sequences that could bind well to L3.Electrophysiological results showed that the three peptides could block obviously the hERG potassium current,showing an effective biological activity,which provides clue for anticancer by targeting hERG potassium channels.

关 键 词：HERG钾通道 噬菌体展示肽库 亲和肽 全自动膜片钳 

分 类 号：R730.2[医药卫生—肿瘤]