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亲环素A抗原表位氨基酸序列及三维结构研究
引用本文:李芳秋,严明,武建国.亲环素A抗原表位氨基酸序列及三维结构研究[J].医学研究生学报,2001,14(3):189-191.
作者姓名:李芳秋  严明  武建国
作者单位:南京军区南京总医院解放军医学检验中心
基金项目:江苏省自然科学基金!资助项目 (BK97174)
摘    要:目的:研究人抗亲环素A(CyPA)McAb D4识别的抗原表位的氨基酸组成及其在蛋白分子上的三维定位。方法:以抗人亲环素A McAb D4作捕获抗体,对噬菌体展示肽库进行淘筛,并对筛选出的噬菌体中插入片段进行DNA序列测定和氨基到序列分析,再用RasMol蛋白质三维结构分析软件在亲环素A分子的三维结构中查找相应的抗原表位。结果:经过三轮淘筛和扩增,选择7个克隆进行序列,提示该抗原表位是构象性的。用RasMol蛋白质三维结构分析软件在亲环素A分子的三维结构中,找到由W121、S99、L98、Q111、S110、F112、L122构成的疏水代状结构,可能就是McAb D4的抗原表位。结论:人亲环素A分子上W121、S99、L98、Q111、S110、F112、L122构成一个构象型抗原表位。

关 键 词:亲环素A  抗原表位  三维结构  噬菌体肽库  亲和淘筛
文章编号:1008-8199(2001)03-0189-03
修稿时间:2000年10月5日

Identification of peptide sequence and three dimensional structure of a epitope of cyclophilin A
LI Fang-qiu,YAN Ming,WU Jian-guo.Identification of peptide sequence and three dimensional structure of a epitope of cyclophilin A[J].Bulletin of Medical Postgraduate,2001,14(3):189-191.
Authors:LI Fang-qiu  YAN Ming  WU Jian-guo
Abstract:Objectives: To identify the peptide sequence of a epitope and study its three dimensional structure on molecule of cyclophilin A. Methods: Using anti oyelophilin A (CyPA) monoclonal antibody (McAb D4) as the capture antibody, the library displayed random heptapeptides was screened by biopanning. The phage clones with high affinity for McAb D4 were chosen and their inserts were sequenced and then the antibody binding site was searched for on three dimensional structure of CyPA with RasMol software. Results:After 3 rounds of biopanning, 7 phage clones with high affinity for McAb D4 were sequenced and 3 different species were obtained, five of seven clones shared a predominant consensus sequence, WSLQSFL. The amino acid sequences of the inserts were compared with that of human CyPA.No homologous sequence was found in the primary structure of CyPA. With the data from PDB database and RasMol software, we found that W121,S99,L98,Q111,S110,F112 and L122 constructed a hydrophobic pocket which may be the epitope for McAb D4. Conclusions: W121,S99,L98,Q111,S110,F112 and L122 may form a conformational epitope of cyclophilin A.
Keywords:Cyclophilin A  Epitope  Phage  display peptide library  Biopanning
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