Structural Biology Core

结构生物学核心

• 批准号: 10508447
• 负责人: Dmitry Lyumkis
• 金额: $ 99.18万
• 依托单位: SEATTLE CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-22 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10508447
• 关键词: 3-Dimensional; Achievement; Address; Affect; Amino Acids; Architecture; Behavior; Binding; Biological; Biology; Chemicals; Collaborations; Collection; Communities; Complex; Consultations; Coupled; Cryo-electron tomography; Cryoelectron Microscopy; Custom; Data; Data Collection; Databases; Deuterium; Development; Educational workshop; Environment; Evaluation; Experimental Designs; Fluorine; Future; Goals; HIV; HIV-1; Hydrogen; Individual; Laboratories; Ligands; Macromolecular Complexes; Mass Spectrum Analysis; Measurement; Methods; Microscopy; Molecular; Molecular Conformation; Molecular Mechanisms of Action; Molecular Probes; Molecular Structure; NMR Spectroscopy; Negative Staining; Nuclear Magnetic Resonance; Nucleic Acid Probes; Nucleic Acids; Operating Rooms; Phase; Play; Proteins; Regulation; Research; Research Personnel; Resolution; Resources; Role; Sampling; Services; Structure; Techniques; Technology; Time; Titan; Tomogram; Training; United States National Institutes of Health; Validation; Viral; Viral Proteins; Virus; Virus Replication; antiretroviral therapy; base; crosslink; design; genetic regulatory protein; image processing; inhibitor therapy; insight; instrumentation; interest; macromolecular assembly; macromolecule; member; new technology; novel; particle; protein complex; protein function; small molecule; structural biology; synergism; tandem mass spectrometry; technology development; tool; voltage

ABSTRACT, CORE 2 Deciphering the molecular mechanisms underlying key steps of the HIV-1 replication cycle is essential to fully describe the biology of the virus, its interplay with host components, and for developing novel antiretroviral therapies. Toward this goal, structural biology plays an important role, as an atomic-level understanding of macromolecular structure and dynamics, and how alterations in structure affect function, provides mechanistic insights into the workings of biological macromolecules. The structural biology core (Core 2) will contribute complementary expertise, instrumentation, and resources for studying key steps of the viral replication cycle, as described within individual projects, including viral entry/uncoating, integration, and maturation/assembly. Core 2 includes: hydrogen deuterium exchange coupled to mass spectrometry (HDX-MS) and cross-linking mass spectrometry (XL-MS), nuclear magnetic resonance (NMR) spectroscopy, single-particle cryo-electron microscopy (cryo-EM), and cryo-electron tomography (cryo-ET). HDX-MS and XL-MS methods have emerged as powerful tools to probe protein interactions with ligands, co-regulatory proteins, and nucleic acids in solution to yield insights into macromolecular behavior on a residue-by-residue level. NMR spectroscopy enables determining high-resolution structures of small-to-medium size proteins and nucleic acids, probing macromolecular interactions at atomic resolution and studying conformational dynamics. Single-particle cryo- EM yields an understanding of macromolecular structure and dynamics for large assemblies, often with less material and more rapidly than with traditional methods. Cryo-ET and sub-tomogram averaging (STA) is ideally suited to studying irregular objects such as non-icosahedral protein lattices, where having 3D volumetric information for each unique assembly, prior to ensemble averaging, is necessary to gain insight into local and long-range structural perturbations. Core 2 will apply existing tools to address specific questions described within individual Projects, and each laboratory will also develop novel technologies to meet the needs of proposed research goals. The Core has established collaborations with most current B-HIVE investigators, has developed workflows for new projects that may emerge during the studies, and will provide ongoing consultation and training to B-HIVE members as projects develop. The established tools provide complementary and synergistic expertise toward the broad research goals of the center, while the training and dissemination module aligns with the broader impacts of the NIH.

摘要，核心2 解解HIV-1复制周期的关键步骤的分子机制对于完全至关重要 描述病毒的生物学，与宿主成分的相互作用，并用于开发新型抗逆转录病毒 疗法。为了实现这一目标，结构生物学起着重要的作用，作为对原子水平的理解 大分子结构和动力学以及结构的变化如何影响功能 洞悉生物大分子的工作。结构生物学核心（核心2）将有助于 互补的专业知识，仪器和资源用于研究病毒复制周期的关键步骤， 在各个项目中描述，包括病毒进入/脱落，集成和成熟/组装。 核心2包括：氢氘交换与质谱（HDX-MS）和交联质量 光谱法（XL-MS），核磁共振（NMR）光谱，单粒子冷冻电子 显微镜（Cryo-EM）和冷冻电子断层扫描（Cryo-ET）。 HDX-MS和XL-MS方法已经出现 作为探测蛋白质与配体，共调节蛋白和核酸中蛋白质相互作用的强大工具 在划分划分水平上对大分子行为的见解。 NMR光谱可以实现 确定小型至中等大小蛋白质和核酸的高分辨率结构，探测 原子分辨率和研究构象动力学的大分子相互作用。单粒子冷冻 EM对大型组合的大分子结构和动力学有了了解，通常很少 物质，比传统方法更快。冷冻-ET和子图平均图（STA）是理想情况下 适合研究不规则的物体，例如非面体蛋白质晶格，其中具有3D体积 在合奏平均之前，每个唯一组件的信息对于洞悉本地和 远程结构扰动。 核心2将应用现有工具来解决各个项目中描述的特定问题，每个工具 实验室还将开发新的技术，以满足拟议的研究目标的需求。核心有 与大多数当前的B-HY研究人员建立了合作，已为新项目开发了工作流程 这可能在研究期间出现，并将为B-HY的成员提供持续的咨询和培训 项目发展。既定的工具为广泛的互补和协同专业知识提供了互补和协同的专业知识 该中心的研究目标，而培训和传播模块与该中心的更大影响一致 NIH。