Structural Biology Core

结构生物学核心

• 批准号: 10650875
• 负责人: Dmitry Lyumkis
• 金额: $ 102.28万
• 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/10650875
• 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; 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 Acids; Operating Rooms; Phase; Play; Proteins; Regulation; Research; Research Personnel; Resolution; Resources; Role; Sampling; Services; Structure; Techniques; Technology; Time; Titan; Training; United States National Institutes of Health; Validation; Viral; Viral Proteins; Virus; Virus Replication; antiretroviral therapy; crosslink; design; 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; videoconference; 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 破译HIV-1复制周期关键步骤的分子机制对于充分研究HIV-1的分子机制至关重要。 描述病毒的生物学，它与宿主成分的相互作用，以及开发新的抗逆转录病毒药物。 治疗为了实现这一目标，结构生物学起着重要的作用，作为一个原子水平的理解， 大分子结构和动力学，以及结构变化如何影响功能，提供了机制 深入了解生物大分子的运作。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， 在单个项目中描述，包括病毒进入/脱壳、整合和成熟/组装。 核心2包括：氢氘交换耦合质谱（HDX-MS）和交联质谱 光谱法（XL-MS），核磁共振（NMR）光谱法，单粒子低温电子 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、共调节蛋白和核酸在溶液中相互作用的有力工具 从而在残基水平上深入了解大分子的行为。核磁共振光谱学使 确定小到中等大小的蛋白质和核酸的高分辨率结构， 原子分辨率的大分子相互作用和研究构象动力学。单粒子低温 EM产生了大分子结构和动力学的大型组件的理解，往往与较少的 材料和更快比传统的方法。冷冻ET和子断层图像平均（STA）是理想的 适用于研究不规则物体，如非二十面体蛋白质晶格，其中具有3D体积 在整体平均之前，每个独特组件的信息对于深入了解局部和 长期结构扰动 核心2将应用现有的工具来解决各个项目中描述的具体问题， 实验室还将开发新技术，以满足拟议研究目标的需要。核心 与大多数当前的B-HIVE研究人员建立了合作关系，为新项目开发了工作流程 研究期间可能出现的问题，并将为B-HIVE成员提供持续的咨询和培训， 项目发展。已建立的工具为广泛的 培训和传播模块与中心的更广泛影响保持一致， 国家卫生研究院