囊泡诱导序列诱导的囊泡蛋白质谱及生物信息学分析  

Protein Profiling and Bioinformatics Analysis of Vesicles Induced by VIS

作　　者：贾万欣 邹聪 张成州 李斌[1,2] 王世民 JIA Wan-xin;ZOU Cong;ZHANG Cheng-zhou;LI Bin;WANG Shi-min(College of Veterinary Medicine,Xinjiang Agricultural University,Urumqi,Xinjiang 830052,China;Xinjiang Key Laboratory of New Drug Study and Creation for Herbivorous Animals,Urumqi,Xinjiang 830052,China)

机构地区：[1]新疆农业大学动物医学学院,新疆乌鲁木齐830052 [2]新疆草食动物新药研究与创制重点实验室,新疆乌鲁木齐830052

出　　处：《动物医学进展》2024年第11期16-22,共7页Progress In Veterinary Medicine

基　　金：新疆维吾尔自治区重点研发计划项目(2023B02012-2);国家自然科学基金项目(31860710)。

摘　　要：囊泡诱导序列(vesicle inducing sequence,VIS)诱导哺乳动物细胞产生的囊泡,能够装载或展示靶蛋白质,具有产量高、易于改造等特征,有望在草食动物疫苗及水产疫苗等领域得到应用。然而,VIS囊泡的结构组成及形成机制目前还不清楚。基于此,将免疫亲和纯化的VIS囊泡进行蛋白质质谱(LC-MS/MS)鉴定,对鉴定结果进行基因本体(gene ontology,GO)功能注释、KEGG代谢通路分析,同时利用Western blot技术验证排名靠前的部分蛋白质。蛋白质质谱技术共鉴定出709种蛋白质。其中,多数蛋白质为细胞囊泡组分,且部分蛋白质参与高尔基体膜泡运输。KEGG通路富集结果显示,VIS囊泡结构蛋白共参与了3条囊泡形成及运输过程相关的代谢通路,分别为吞噬体通路、内吞作用、突触囊泡途径。该研究从蛋白质水平上揭示了VIS囊泡构成及形成机制,对今后VIS囊泡在动物生物制品及疫苗等领域的应用奠定了基础。The vesicle-inducing sequence(VIS)induces the formation of vesicles in mammalian cells,which can load or display target proteins.VIS vesicles exhibit characteristics such as high yield and ease of modification,making them promising for applications in the fields of herbivorous animal vaccines and aquatic vaccines.However,the structural composition and formation mechanism of VIS vesicles are currently unclear.In this study,immunopurified VIS vesicles were subjected to protein identification using liquid chromatography-tandem mass spectrometry(LC-MS/MS).The identified proteins were annotated for Gene Ontology(GO)functionality,and KEGG metabolic pathway analysis was performed.Additionally,Western blot techniques were employed to validate selected proteins from the identification results.A total of 709 proteins were co-identified using proteomic techniques.Among them,the majority were cellular vesicle components,with some proteins participating in the transport of Golgi membrane vesicles.The KEGG pathway enrichment analysis revealed that the identified proteins in VIS vesicles were involved in three metabolic pathways related to vesicle formation and transport:autophagy,endocytosis,and synaptic vesicle pathways.This study,through a proteomic approach,elucidates the composition and formation mechanism of VIS vesicles,laying a foundation for the future application of VIS vesicles in the fields of animal bioproducts and vaccines.

关 键 词：囊泡诱导序列 囊泡 免疫亲和 质谱鉴定 高尔基体 

分 类 号：S852.5[农业科学—基础兽医学]