人67 LR噬菌体结合肽的筛选与功能分析  

Screening and Functional Analysis of Human 67LR-binding Phage Display Peptides

作　　者：连继勤[1] 戴旭芳[2] 卢忠燕[1] 王喆[1] 甘立霞[1] 何凤田[1] 

机构地区：[1]第三军医大学生物化学与分子生物学教研室,重庆市400038 [2]重庆师范大学特殊教育学院,重庆市400047

出　　处：《医学分子生物学杂志》2008年第6期476-479,共4页Journal of Medical Molecular Biology

基　　金：国家自然科学基金(No.30400559);重庆市自然科学基金(No.2008BB5112)~~

摘　　要：目的为了获得能够与人67ku层黏连蛋白受体(67 ku laminin receptor,67LR)特异性结合的噬菌体呈现肽。方法以重组人67LR蛋白为诱饵,分别对噬菌体12肽库和环7肽库进行亲和凝胶筛选,ELISA测定所筛选噬菌体克隆与靶蛋白的亲和力,经测序分析得到67LR特异性结合噬菌体克隆,并通过293细胞侵袭转移实验测定阳性噬菌体呈现肽对67LR促细胞侵袭作用的抑制效果。结果经4轮筛选共筛到2类新的67LR结合肽,分别具有DXCETCT(X可变)和YRPMXEY(X可变)一致序列。其中DX-CETCT噬菌体呈现肽可显著抑制67LR所诱导的细胞侵袭,且这种抑制作用与YIGSR5肽具有一定互补性。结论成功筛选到2类新的67LR结合肽,其中DXCETCT噬菌体呈现肽可针对性抑制67LR所诱导的细胞侵袭作用。Objective To obtain functional phage display peptides specifically binding to human 67ku laminin receptor （67LR） . Methods Phage random 7- and 12-peptide libraries were screened using recombinant 67LR protein as a bait. ELISA and sequencing were used to identify positive phages. Invasion assay in vitro were used to analysis the function of positive phage display peptides. Results After 4 rounds of panning, total 2 types of new peptides binding to 67LR specifically were screened, with DXCETCT （X is alterable） and YRPMXEY （X is alterable） consensus sequences respectively. The invasion of 293 cell induced by 67LR was suppressed by DXCETCT display peptides, which was complementarily strengthened by YIGSR peptide. Conclusion The peptide DXCETCT with an alterable amino acid at the position of X is a new type of 67LR suppressing peptide.

关 键 词：噬菌体呈现肽 层黏连蛋白受体 

分 类 号：R782[医药卫生—口腔医学]