脂多糖结合蛋白与脂多糖结合位点模拟肽的筛选  

Screening of mimic peptides of lipopolysaccharide binding protein binding to LPS

作　　者：杨海捷[1] 杨韶艳[1] 刘晓丽[1] 谭志胜[1] 郭军[1] 王红[1] 

机构地区：[1]成都军区昆明总医院军队干部病房,昆明650032

出　　处：《中华保健医学杂志》2013年第5期415-418,共4页Chinese Journal of Health Care and Medicine

基　　金：国家自然基金课题(81270224)

摘　　要：目的应用噬菌体随机12肽库筛选脂多糖结合蛋白(LBP)与脂多糖结合位点的多肽序列。方法以脂多糖为固相筛选分子,对噬菌体12肽库进行4轮亲合筛选,采用酶联免疫吸附法鉴定筛选后的噬菌体克隆与脂多糖的结合活性,取结合活性较高的克隆进行DNA测序,推导其多肽序列并与LBP序列比对,再进行竞争抑制实验检测筛选噬菌体与LBP竞争结合的效力。结果选取的86个噬菌体克隆中,多数结合实验鉴定阳性,取20个亲和力最高的克隆测序,得到20条不同编码的12肽序列,20条多肽与LBP一级结构有不同程度同源性,均为模拟肽。12个噬菌体克隆有较高的竞争抑制率。结论用噬菌体12肽库成功筛选出LBP与脂多糖结合位点模拟肽,为进一步以这些多肽为先导物进行定向进化研究提供了实验依据和结构基础。Objective To screen the mimic peptides of lipopolysaccharide binding protein （LBP） binding to LPS from phage display random peptide library. Methods Using LPS as a target molecule,4 rounds of biopanning to a phage random 12-mer peptide library were carried out. Twenty clones were selected randomly and used in binding test. Positive clones with high affinity were sequenced,and these peptides were compared with LBP sequence. Competitive inhibition test was carried out. Results Most Out of the 86 clones were identified as positive. 20 of the 86 had higher affinity,and encoded 20 different 12-mer peptides which were not homologous with the sequence of LBP. The phage clones with these peptides had high competitive inhibition with LBP. Conclusions Mimic peptides of LBP binding to LPS are obtained by phage peptide library screening, which provide us some new laboratory evidence and structural foundations on further study of direct-evolution using these peptides as primers.

关 键 词：脂多糖结合蛋白 噬菌体肽库 模拟肽 

分 类 号：R34[医药卫生—基础医学]