拟穴青蟹细丝蛋白C的生物信息学分析及重组表达  

Bioinformatics Analysis and Recombinant Expression of Filamin C in Scylla paramamosain

作　　者：何欣蓉 杨阳 张永霞 刘红 曹敏杰 刘光明 HE Xinrong;YANG Yang;ZHANG Yongxia;LIU Hong;CAO Minjie;LIU Guangming(College of Food and Biological Engineering,Jimei University,Xiamen 361021,China;National & Local Joint Engineering Research Center of Processing Technology for Aquatic Products,Xiamen 361021,China;Xiamen Key Laboratory of Marine Functional Food,Xiamen 361021,China)

机构地区：[1]集美大学食品与生物工程学院,福建厦门361021 [2]水产品深加工技术国家地方联合工程研究中心,福建厦门361021 [3]厦门市海洋功能食品重点实验室,福建厦门361021

出　　处：《集美大学学报（自然科学版）》2021年第1期14-21,共8页Journal of Jimei University：Natural Science

基　　金：国家自然科学基金项目(31871720,U1812403);福建省科技计划项目(2018N5009,2018R0071);海洋科研专项基金项目(DY135-B2-07)。

摘　　要：为了探讨抗原表位与细丝蛋白C(filamin C,FLN c)结构之间的构效关系,对FLN c进行生物信息学分析及分段重组表达。生物信息学分析结果表明,FLN c二级结构以β折叠及无规则卷曲为主,确定其具有9个结构域,三级结构呈免疫球蛋白样折叠,具有典型的细丝蛋白家族特征。基于生物信息学分析,将拟穴青蟹(Scylla paramamosain)FLN c通过分段原核表达,纯化出带有麦芽糖结合蛋白(maltose binding protein,MBP)标签的重组表达蛋白rFLN c1(AA:1-335)、rFLN c2(AA:336-531)、rFLN c3(AA:532-847)。血清学分析结果显示,rFLN c2与蟹类过敏患者血清IgE结合能力最强,推测此区域含有大部分抗原表位。因此,定位拟穴青蟹FLN c氨基酸336-531结构域为抗原表位优势区,rFLN c2可用于FLN c的晶体学研究。In order to explore the structure-activity relationship between the epitope and the structure of filamin C(FLN c),bioinformatics analysis and segmented recombinant expression of FLN c were performed.The results of bioinformatics analysis showed that the secondary structure of FLN c had 9 domains mainly withβ-sheet and random coil.The tertiary structure was an immunoglobulin-like fold and had a typical filamin family characteristics.Based on bioinformatics analysis,the fragmented FLN c were expressed in Escherichia coli,and the recombinant protein rFLN c1(AA:1-335),rFLN c2(AA:336-531)and rFLN c3(AA:532-847)with maltose binding protein tag were purified.The results of serological analysis showed that rFLN c2 had the strongest binding ability to IgE in sera of crab allergy patients.It was speculated that this region contained most of the epitopes.So the amino acid 336-531 domain of Scylla paramamosain FLN c as the epitope predominant region was mapped,indicating that rFLN c2 could be used for crystallographic research of FLN c.

关 键 词：拟穴青蟹 细丝蛋白C 生物信息学 重组表达 

分 类 号：Q816[生物学—生物工程]