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：艾滋病毒附件和调节复合物 蛋白质组学核心 总结 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 基于质谱（MS）的蛋白质组学技术用于HIV宿主蛋白的功能和结构表征 配合物与遗传学，结构生物学和计算核心一起，我们提供了一个 用于Vif-、达特-和Rev-宿主蛋白质复合物的结构测定的系统管道。这本小说 HARC内源性蛋白质结构（HEPS）平台将收集和整合结构数据， 内源性纯化病毒-宿主复合物的综合结构模型。蛋白质组学核心将 进行内源性蛋白质纯化，并绘制HIV和宿主蛋白质的蛋白质-蛋白质相互作用（PPI）图谱 在HIV感染的和基因组编辑和未经编辑的原代CD 4 + T细胞中，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， 氢/氘交换-MS（H/DX-MS），以支持具有挑战性和灵活性的 HIV-宿主蛋白复合物。最后，为了表征参与细胞凋亡的染色质和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）和基因组修饰的CD 4+原代细胞感染WT或突变型HIV。