Core 1 Krogan

HARC中心：HIV附件和调控复合体 蛋白质组学核心 摘要 蛋白质组学核心将通过应用和开发质谱学来支持所有HARC中心项目 基于(MS)的蛋白质组学技术用于HIV宿主蛋白的功能和结构表征 复合体。与遗传学、结构生物学和计算核心一起，我们提供了 Vif-、Tat-和Rev-宿主蛋白复合体结构测定的系统流水线。这部小说 HARC内源蛋白质结构(HEPS)平台将收集和整合结构数据，以构建 内源性纯化的病毒-宿主复合体的综合结构模型。蛋白质组学的核心将 执行内源性蛋白质提纯并绘制HIV和宿主蛋白质的蛋白质-蛋白质相互作用图(PPI) 在HIV感染和通过亲和纯化进行基因组编辑和未编辑的原代CD4+T细胞中，我们将 此外，还提供结构蛋白质组学方法，包括天然MS(NM)、交联型(XL)-MS和 氢/氢交换-质谱仪(H/DX-MS)，支持结构测定的挑战性和灵活性 HIV-宿主蛋白复合体。最后，为了表征染色质和RNA结合蛋白的变化， HIV转录和潜伏期，我们将识别和定量比较翻译后修饰(PTM) 野生型(WT)和基因修饰的CD4+原代细胞感染野生型(WT)或突变HIV。

THE HARC CENTER: HIV ACCESSORY AND REGULATORY COMPLEXES PROTEOMICS CORE SUMMARY The Proteomics Core will support all HARC Center Projects by applying and developing mass spectrometry (MS)-based proteomic technologies for the functional and structural characterization of HIV-host protein complexes. Together with the Genetics, Structural Biology, and Computational Cores, we provide a systematic pipeline for the structure determination of Vif-, Tat-, and Rev-host protein complexes. This novel HARC endogenous protein structure (HEPS) platform will collect and integrate structural data to build comprehensive structural models of endogenously purified virus-host complexes. The Proteomics Core will perform endogenous protein purifications and map protein-protein interactions (PPIs) of HIV and host proteins in HIV-infected and genomically edited and unedited primary CD4+ T cells by affinity-purification MS. We will additionally provide structural proteomics methodologies, including native MS (nM); cross-linking (XL)-MS, and hydrogen/deuterium exchange-MS (H/DX-MS), to support the structure determination of challenging and flexible HIV-host protein complexes. Finally, to characterize changes to chromatin and RNA-binding proteins involved in HIV transcription and latency, we will identify and quantitatively compare post-translational modifications (PTMs) of wild-type (WT) and genomically modified CD4+ primary cells infected with WT or mutant HIV.