Structural Biology Core

结构生物学核心

基本信息

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
项目类别：
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

项目摘要

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 复制周期关键步骤背后的分子机制对于充分了解 HIV-1 复制周期至关重要描述病毒的生物学、其与宿主成分的相互作用，以及开发新型抗逆转录病毒药物疗法。为了实现这一目标，结构生物学发挥着重要作用，作为对原子水平的理解大分子结构和动力学，以及结构的改变如何影响功能，提供了机制深入了解生物大分子的工作原理。结构生物学核心（核心2）将做出贡献用于研究病毒复制周期关键步骤的互补专业知识、仪器和资源，例如在各个项目中进行了描述，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) 和交联质量光谱测定 (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溶液中蛋白质与配体、共调控蛋白质和核酸相互作用的强大工具深入了解逐个残留物水平上的大分子行为。核磁共振波谱能够确定中小型蛋白质和核酸的高分辨率结构，探测原子分辨率下的大分子相互作用和研究构象动力学。单粒子冷冻 EM 可以帮助您了解大型组装体的大分子结构和动力学，通常只需较少的时间材料，并且比传统方法更快。冷冻电子断层扫描 (Cryo-ET) 和亚断层扫描平均 (STA) 是理想的选择适合研究不规则物体，例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本地和长程结构扰动。核心 2 将应用现有工具来解决各个项目中描述的具体问题，并且每个项目实验室还将开发新技术以满足拟议研究目标的需要。核心有与最新的 B-HIVE 研究人员建立了合作，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开发。已建立的工具为广泛的领域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广泛影响相一致 NIH。