间隙连接蛋白Cx31.1相互作用蛋白的初步鉴定

Protein Interaction between Connexin 31.1 and Proteins Related to Reproduction

目的:筛选间隙连接蛋白31.1(Cx31.1)的相互作用蛋白,进一步阐释Cx31.1在生殖健康方面可能的作用机制。方法:采用免疫共沉淀技术富集稳定表达Cx31.1-EGFP融合蛋白细胞中的Cx31.1结合蛋白,以稳定表达EGFP标签蛋白的细胞作为对照组。得到的蛋白复合物直接酶解后进行高效液相色谱-串级质谱(HPLC-MS/MS)分析,通过相应的肽段序列标签在Swiss Prot数据库中搜索蛋白质。结果:通过数据库搜索首次鉴定了5个与生殖相关的且与Cx31.1可能发生相互作用的蛋白,分别是HSPB1、HSPA2、HSPA5、BYP2和XAGE1A。其中HSPB1、HSPA2和HSPA5属于热休克蛋白家族,BYP2和XAGE1A蛋白在睾丸组织中特异表达。结论:Cx31.1可能与生殖相关蛋白具有相互作用。

Objective：To identify connexin 31.1（Cx31.1） interacting proteins and find out possible mechanism of Cx31.1 in the function of reproduction.Methods：Cx31.1 interacting proteins were separated from cells stably expressing Cx31.1-EGFP using immunoprecipitation,and proteins separated from cells stably expressing EGFP were used as the control.After in-gel digested and analyzed by HPLC-MS/MS,the possible interacting proteins were identified by peptide sequence tags and database searching in Swiss-prot.Results：Five Cx31.1 binding proteins were firstly identified by database searching,which were HSPB1,HSPA2,HSPA5,BYP2 and XAGE1A.HSPB1,HSPA2 and HSPA5 belong to heat shock protein,while BYP2 and XAGE1A belong to the family of GAGE were specifically expressed in testis.Conclusion：Cx31.1 is a protein involved in reproduction by interacting with some reproduction-related proteins.