Core 1 Krogan

核心 1 克洛根

• 批准号: 10506983
• 负责人: Danielle L Swaney
• 金额: $ 50.07万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-15 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10506983
• 关键词: Acetylation; Affinity Chromatography; Biological Assay; CD4 Positive T Lymphocytes; CRISPR/Cas technology; Cell Line; Cell model; Cells; Chromatin; Collaborations; Complex; Consequences of HIV; Cryoelectron Microscopy; Data; Dependence; Deuterium; Foundations; Gene Deletion; Genetic; Genetic Transcription; Genomics; Goals; HIV; Hydrogen; In Vitro; Individual; Integration Host Factors; Knock-out; Maps; Mass Spectrum Analysis; Measures; Membrane Proteins; Methodology; Modeling; Phosphorylation; Point Mutation; Post-Translational Protein Processing; Process; Protein-Protein Interaction Map; Proteins; Proteomics; RNA-Binding Proteins; Rest; Services; Signal Pathway; Solvents; Structural Models; Structure; Surface; System; Technology; Tertiary Protein Structure; Ubiquitination; Viral Proteins; Virus; Work; base; crosslink; experimental study; flexibility; innovation; interest; mutant; novel; protein complex; protein protein interaction; protein purification; protein structure; stoichiometry; structural biology

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.

HARC 中心：HIV 配件和调节复合体 蛋白质组学核心 概括 蛋白质组学核心将通过应用和开发质谱来支持所有 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 复合物。与遗传学、结构生物学和计算核心一起，我们提供了 用于确定 Vif-、Tat- 和 Rev- 宿主蛋白复合物结构的系统流程。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来构建 内源纯化的病毒-宿主复合物的综合结构模型。蛋白质组学核心将 进行内源蛋白纯化并绘制 HIV 和宿主蛋白的蛋白间相互作用 (PPI) 通过亲和纯化 MS 检测 HIV 感染的、经过基因组编辑和未经编辑的原代 CD4+ T 细胞。我们将 另外提供结构蛋白质组学方法，包括天然 MS (nM)；交联 (XL)-MS，和 氢/氘交换-MS (H/DX-MS)，支持具有挑战性且灵活的结构测定 HIV-宿主蛋白复合物。最后，为了表征参与染色质和 RNA 结合蛋白的变化 HIV 转录和潜伏期，我们将识别并定量比较翻译后修饰 (PTM) 感染 WT 或突变型 HIV 的野生型 (WT) 和基因修饰 CD4+ 原代细胞。