家蚕微孢子虫孢原质与家蚕细胞表面互作蛋白质组的鉴定分析  

Identification of Surface Proteins Mediating Interactions between Nosema bombycis Sporoplasm and Silkworm Cell

作　　者：何雪梅[1,2] 何强[1,2] 韦俊宏[1,2] 李春峰[1,2] 潘国庆[1,2] 周泽扬[1,2,3] 李田[1,2] HE Xuemei;HE Qiang;WEI Junhong;LI Chunfeng;PAN Guoqing;ZHOU Zeyang;LI Tian(State Key Laboratory of Resource Insects,Southwest University,Chongqing 400715,China;Chongqing Key Laboratory of Microsporidia Infection and Control,Southwest University,Chongqing 400715,China;College of Life Science,Chongqing Normal University,Chongqing 401331,China)

机构地区：[1]资源昆虫高效养殖与利用全国重点实验室(西南大学),重庆400715 [2]西南大学微孢子虫感染与防控重庆市重点实验室,重庆400715 [3]重庆师范大学生命科学学院,重庆401331

出　　处：《蚕学通讯》2024年第4期68-76,共9页Newsletter of Sericultural Science

基　　金：国家自然科学基金面上项目(32272944);重庆市自然科学基金创新群体项目(cstc2021jcyj-cxtt0005);中央高校基本科研业务费项目(SWU-XDJH202322)。

摘　　要：微孢子虫(Microsporidia)是一类专性细胞内寄生的单细胞真核微生物,通过极管弹出孢原质的方式进行感染,因此孢原质是微孢子虫完成感染的重要中间形态,但孢原质与宿主细胞的相互作用尚缺乏深入研究。本研究利用生物素-链霉亲和素系统,对介导家蚕微孢子虫(Nosema bombycis)孢原质与家蚕胚胎细胞(BmE)粘附的表面蛋白质组进行了分离和质谱鉴定分析,获得了32个家蚕微孢子虫孢原质表面蛋白和188个家蚕细胞表面蛋白。对蛋白质进行功能和通路富集分析发现,所鉴定的家蚕细胞表面互作蛋白主要参与膜物质运输,可能在介导孢原质侵入过程中发挥作用。通过对蛋白质进行亚细胞定位分析,筛选了6个家蚕微孢子虫蛋白和10个家蚕细胞表面蛋白作为候选互作分子。利用酵母双杂交实验对候选互作蛋白进行验证分析发现,家蚕微孢子虫的Nb1BD、Nb4BD和Nb6BD与家蚕的Bm4AD和Bm6AD之间存在相互作用关系。本研究首次获得了介导孢原质与宿主细胞互作的表面蛋白,为深入解析家蚕微孢子虫的侵染机制提供了重要参考数据。Microsporidia are a group of obligate intracellular and unicellular eukaryotic parasites that infect hosts through the ejection of sporoplasm via polar tubules,so sporoplasm is an important intermediate form in microsporidia completion of infection.However,the interaction between sporoplasm and host cells lacks in-depth research.In this study,we used the biotin-streptavidin system to isolate and perform mass spectrometry-based identification of surface proteins mediating the adhesion of Nosema bombycis sporoplasm to Bombyx mori BmE cells.We identified 32 surface proteins of N.bombycis sporoplasm and 188 surface proteins of BmE cells.Function and pathway enrichment analyses of the identified proteins revealed that the B.mori proteins mainly participate in membrane material transport and may play a role in mediating sporoplasm invasion.Subcellular localization analysis identified 6 N.bombycis proteins and 10 B.mori proteins as candidate interacting molecules.Yeast two-hybrid experiments were used to verify the interaction between candidate interacting proteins,and we found that Nb1BD,Nb4BD,and Nb6BD of N.bombycis interact with Bm4AD and Bm6AD of B.mori.This study has identified surface proteins mediating adhesion between sporoplasm and host cells,providing important reference data for further elucidating the infection mechanism of N.bombycis.

关 键 词：家蚕 家蚕微孢子虫 孢原质 家蚕胚胎细胞 表面互作蛋白 

分 类 号：R73[医药卫生—肿瘤]