鼠伤寒沙门氏菌SptP蛋白与RAW264.7细胞互作蛋白的筛选及分析  

Screening and Analysis of Salmonella typhimurium SptP Proteins Interacting with RAW264.7 Cells

作　　者：丁永辉 周南龙 何婷 李天森 DING Yong-hui;ZHOU Nan-long;HE Ting;LI Tian-sen(School of Tropical Agriculture and Forestry,Hainan University,Haikou,Hainan 570228,China)

机构地区：[1]海南大学热带农林学院,海南海口570228

出　　处：《动物医学进展》2024年第11期22-28,共7页Progress In Veterinary Medicine

基　　金：海南省基础与应用基础研究计划高层次人才项目(2019RC021);海南省自然科学基金高层次人才项目(823RC468);海南省研究生创新科研课题(Qhys2021-161)。

摘　　要：为了确定宿主与鼠伤寒沙门氏菌关键毒力蛋白SptP相互作用的蛋白,为鼠伤寒沙门氏菌的防治提供理论依据。通过诱导可溶性His-SptP融合蛋白表达,进行His pull-down试验筛选RAW264.7与SptP互作的蛋白,并经SDS-PAGE及质谱验证,最后通过生物信息学工具分析潜在互作蛋白。结果表明,可溶性His-SptP成功诱导表达;经His pull-down及质谱分析,鉴定到SptP与RAW264.7的153种潜在互作蛋白。另根据GO富集分析,这些互作蛋白主要集中于细胞构造、细胞内部结构和细胞器等,大部分为结合(binding)蛋白质,主要参与细胞过程、代谢过程和有机物代谢过程等。KEGG通路富集结果表明,互作的蛋白质主要参与聚集的通路为细菌对上皮细胞的侵袭、肌动蛋白细胞骨架的调节及沙门氏菌感染等。综上所述,在RAW264.7细胞中筛选到153个与SptP潜在互作的蛋白,这些蛋白在细菌侵入过程中发挥着重要的作用,为SptP在入侵宿主细胞过程中的具体的作用机制的研究提供了一定的理论依据。The purpose of this study is to identify the proteins that interact with the key virulence protein SptP of Salmonella typhimurium and provide the theoretical basis for the prevention and treatment of Salmonella typhimurium.By inducing the expression of soluble His-SptP fusion protein,the protein interacting with RAW264.7 and SptP was screened by His pull-down,which were verified by SDS-PAGE and mass spectrometry.Finally,the potential interacting proteins were analyzed by bioinformatics tools.The results showed that soluble His-SptP was successfully induced and expressed.153 potential interaction proteins between SptP and RAW264.7 were identified by His pull-down and mass spectrometry analysis.According to GO enrichment analysis,these interacting proteins are mainly concentrated in cell structure,cell internal structure,and organelles,and most of them are binding proteins,which mainly participate in cell process,metabolism process,and organic matter metabolism process.The enrichment results of the KEGG pathway showed that the main pathways involved in the aggregation of proteins were bacterial invasion of epithelial cells,actin cytoskeleton regulation,and Salmonella infection.To sum up,153 proteins with potential interaction with SptP were screened in RAW264.7 cells,which played an important role in the process of bacterial invasion,providing a certain theoretical basis for the study of the specific mechanism of SptP in the process of invasion into host cells.

关 键 词：鼠伤寒沙门氏菌 SptP 蛋白互作 巨噬细胞 

分 类 号：S855.1[农业科学—临床兽医学] S858.91[农业科学—兽医学]