兼职蛋白热休克蛋白70与α-烯醇酶的人B细胞表位肽预测及其瓜氨酸化的血清学检定  

Epitopes prediction of moonlighting proteins and determination of the specific autoantibodies in human serum

作　　者：李虹颖 张鑫[1] 王文静 李陈铎[1] 夏思思 吕喆 陈彦[1] 孙英[1] 袁慧慧[1] 王炜[1] LI Hong-ying;ZHANG Xin;WANG Wen-jing;LI Chen-duo;XIA Si-si;Lü Zhe;CHEN Yan;SUN Ying;YUAN Hui-hui;WANG Wei(Department of Immunology,School of Basic Medical Sciences,Capital Medical University,Beijing 100069,China;不详)

机构地区：[1]首都医科大学基础医学院免疫学系,北京100069 [2]首都医科大学附属佑安医院临床检验中心,北京100069 [3]首都医科大学附属北京朝阳医院风湿免疫科,北京100020

出　　处：《微生物学免疫学进展》2021年第3期5-13,共9页Progress In Microbiology and Immunology

基　　金：国家自然科学基金项目(82090013,82071805);北京市自然科学基金项目(7192023)。

摘　　要：目的筛选与自身免疫病相关兼职蛋白的B细胞优势表位肽并进行血清学检定。方法利用DNAMAN软件分析不同物种热休克蛋白70(heat shock protein 70, HSP70)和α-烯醇酶氨基酸序列同源性,IEDB数据库和瓜氨酸化在线平台预测线性B细胞表位和瓜氨酸化修饰位点;间接ELISA检测抗HSP70和α-烯醇酶蛋白瓜氨酸化前后表位肽抗体。结果同源性分析和IEDB数据库预测得到兼职蛋白HSP70和α-烯醇酶各10个B细胞优势表位肽;瓜氨酸化位点预测显示,其分别有9个和4个位点可能会发生瓜氨酸化修饰,其中5个瓜氨酸化修饰位点与预测表位肽重合。与正常人相比,抗环瓜氨酸肽抗体阳性患者血清抗瓜氨酸化修饰表位肽的抗体滴度明显增高(P<0.05)。结论采用生物信息学分析和ELISA初步验证的方法有助于筛选自身抗原相关的B细胞优势表位肽。Objective The present study aims to provide preliminary data for screening the dominant epitopes of antigens recognized by B cells which associate with autoimmune diseases, and to detect them by ELISA. Methods DNAMAN software was employed to analyze the amino acid sequence homology of HSP70 and α-enolase from different species. Then, IEDB database and the online platform of citrullination were used to predict the possible linear B cell epitopes and citrullination sites, respectively. Following that, the indirect ELISA method was used to detect antibodies against the HSP70 and α-enolase protein synthesized peptides prior and post citrullination in serum. Results Based on the data of DNAMAN software and IEDB database, 20 epitope peptides of the human HSP70 and α-enolase proteins(10 for each) were obtained, in which there were nine and four potential sites that might undergo citrullination modification. Furthermore, there were 5 sites of citrullination modification within in the predicted linear B-cell dominant epitope peptides. ELISA data revealed that comparing with control subjects, serums of CCP-positive subjects contained antibodies specifically against the predicted epitope peptides, while the titers of antibodies against the citrullinated epitope peptides were significantly elevated(P<0.05). Conclusion Usage of the methods combined bioinformatics analysis with ELISA can help to provide preliminary data for screening the dominant epitopes recognized by B cells.

关 键 词：兼职蛋白 热休克蛋白 烯醇化酶 瓜氨酸化 优势表位 

分 类 号：R392[医药卫生—免疫学]