细胞外囊泡 (EV) 通过蛋白质、核酸和脂质货物影响细胞表型和功能。由于人们对不同的生物发生机制仍知之甚少，电动汽车具有异质性。我们之前的研究表明，内质网 (ER) 膜接触位点 (MCS) 连接蛋白囊泡相关蛋白相关蛋白 A (VAP-A) 驱动富含 RNA 的 EV 子集的生物发生。在这里，我们检查了 VAP-A 调节的 EV 的蛋白质含量。使用无标记蛋白质组学，我们鉴定了从 VAP-A 敲低 (KD) 结肠癌细胞中纯化的小 EV (SEV) 中下调和上调的蛋白质。数据的基因集富集分析 (GSEA) 揭示了根据 VAP-A 差异分类为 SEV 的蛋白质类别。互惠基因检索工具 (STRING) 对 RNA 结合蛋白 (RBP) 基因集的蛋白质-蛋白质相互作用网络分析发现了几种在 VAP-A KD SEV 中下调的 RNA 功能机制，包括核糖体、剪接体、mRNA 监测和 RNA 转运蛋白。我们还观察到其他功能性相互作用蛋白网络的下调，包括钙粘蛋白结合、未折叠蛋白结合和 ATP 依赖性蛋白。© 2023 Wiley-VCH GmbH。

Extracellular vesicles (EVs) influence cell phenotypes and functions via protein, nucleic acid, and lipid cargoes. EVs are heterogeneous, due to diverse biogenesis mechanisms that remain poorly understood. Our previous study revealed that the endoplasmic reticulum (ER) membrane contact site (MCS) linker protein vesicle associated protein associated protein A (VAP-A) drives biogenesis of a subset of RNA-enriched EVs. Here, we examine the protein content of VAP-A-regulated EVs. Using label-free proteomics, we identified down- and upregulated proteins in small EVs (SEVs) purified from VAP-A knockdown (KD) colon cancer cells. Gene set enrichment analysis (GSEA) of the data revealed protein classes that are differentially sorted to SEVs dependent on VAP-A. Search Tool for the Retrieval of Reciprocity Genes (STRING) protein-protein interaction network analysis of the RNA-binding protein (RBP) gene set identified several RNA functional machineries that are downregulated in VAP-A KD SEVs, including ribosome, spliceosome, mRNA surveillance, and RNA transport proteins. We also observed downregulation of other functionally interacting protein networks, including cadherin-binding, unfolded protein binding, and ATP-dependent proteins.© 2023 Wiley-VCH GmbH.