MicroCal PEAQ-DSC

MicroCal PEAQ-DSC

• 批准号: 10047566
• 负责人: ANGELA M. GRONENBORN
• 金额: $ 13.25万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-11-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10047566
• 关键词: Acquired Immunodeficiency Syndrome; Administrative Supplement; Binding; Biochemical; Biophysics; Capsid; Cell Surface Receptors; Cells; Complex; Computer Analysis; DNA Repair; Disease; Event; Funding; HIV; HIV-1; Immunologic Factors; Individual; Infection; Intervention; Life Cycle Stages; Membrane Fusion; Nuclear; Predisposition; Process; Production; Proteins; Proteomics; Research; Resolution; Reverse Transcription; Scanning; Structure; Time; Viral; Viral Genome; Virus; Virus Diseases; Virus Integration; fight against; imaging study; instrumentation; new therapeutic target; novel; novel therapeutic intervention; programs; protein complex; protein function; success; virology

PCHPI Project Summary Essentially every step in the HIV life cycle interfaces intimately with the host cell machinery. The Pittsburgh Center of HIV Protein Interactions focuses on the steps and interactions that occur with the host after engagement of cell surface receptors and membrane fusion through integration of the viral genome into that of the host, the so called “early events”. Several essential molecular interactions and enzymatic activities occur within this time window, necessary for productive progression of the viral life cycle. Thus, it represents a pivotal period in the infection process, during which the susceptibility of the virus to disruptive interventions is likely to be high and little explored. Broadly speaking, the processes that we focus on include uncoating (the process by which the capsid disassembles), reverse transcription, evasion from innate immune factors, nuclear entry and integration. Given the importance of the capsid structure and its interactions for many of these processes, we also explore maturation and in particular formation of the capsid core. We are building on our successes and applying the extensive and complementary experimental expertise of our team to carry out 1) biochemical and high-resolution structure studies of individual proteins and complexes, 2) biochemical, biophysical, and proteomics analyses to identify novel interactions and complexes, 3) virology and imaging studies to understand protein function in the context of the cell and virus infection, and 4) computational analyses to elucidate the physical basis of capsid formation and capsid interactions with binding partners. Broadly, the program comprises projects on capsid interactions, the engagement of Vpr with the DNA repair machinery, and retroviral intasome structure. The requested administrative supplement will provide funds for the purchase of a differential scanning calorimeter, instrumentation essential for continued rigor in our protein production and characterization pipeline toward high-resolution structures of HIV-host protein complexes. Relevance Results provided by the proposed research are expected to have major implications in the global fight against AIDS, still considered an incurable disease with a pressing need for new therapeutic strategies and novel drug targets. Identifying and characterizing atomic structures of key HIV-1 host protein interactions in the immediate post-entry stage of the virus lifecycle will open new avenues in this endeavor.

PCHPI项目总结 基本上，艾滋病毒生命周期的每一步都与宿主细胞机制密切相关。匹兹堡 HIV蛋白质相互作用中心专注于HIV感染后与宿主发生的步骤和相互作用。 通过病毒基因组整合到细胞表面受体和膜融合的参与， 主持人，所谓的“早期事件”。几个基本的分子相互作用和酶活性发生 在这个时间窗口内，病毒生命周期的生产性进展所必需的。因此，它代表一个 在感染过程中的关键时期，在此期间，病毒对破坏性干预的敏感性是 可能很高，而且很少有人探索。一般来说，我们关注的过程包括去涂层（ 衣壳解体的过程），逆转录，逃避先天免疫因子，核 进入和融合。考虑到衣壳结构及其相互作用对许多这些病毒的重要性， 过程中，我们还探讨了成熟，特别是形成的衣壳核心。我们正在建立我们的 成功和应用我们的团队的广泛和互补的实验专业知识，以执行1） 单个蛋白质和复合物的生物化学和高分辨率结构研究，2）生物化学， 生物物理学和蛋白质组学分析，以确定新的相互作用和复合物，3）病毒学和成像 研究以了解细胞和病毒感染背景下的蛋白质功能，以及4）计算 分析以阐明衣壳形成和衣壳与结合配偶体相互作用的物理基础。 广义上，该计划包括衣壳相互作用的项目，Vpr与DNA修复的参与 机器和逆转录病毒intasome结构。所要求的行政补充经费将用于 购买差示扫描量热仪，这是我们蛋白质持续严谨所必需的仪器 生产和表征管道，以高分辨率的HIV-宿主蛋白质复合物的结构。 相关性 拟议研究提供的结果预计将对全球防治艾滋病产生重大影响。 艾滋病仍然被认为是一种不治之症，迫切需要新的治疗策略和新药 目标的识别和表征关键HIV-1宿主蛋白相互作用的原子结构 病毒生命周期的进入后立即阶段将为这一奋进开辟新的途径。