Capturing structure and dynamics of transmembrane signaling proteins

捕获跨膜信号蛋白的结构和动态

• 批准号: 10367643
• 负责人: Taras V. Pogorelov
• 金额: $ 30.61万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-20 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10367643
• 关键词: Binding; Biology; Cell Signaling Process; Cell membrane; Cells; Cellular Membrane; Characteristics; Chemicals; Clinical; Communities; Competence; Complex; Computer software; Computing Methodologies; Data; Development; Diabetes Mellitus; Dimerization; Disease; Entropy; Environment; Extracellular Domain; FGFR3 gene; Family; Fluorescence Resonance Energy Transfer; Free Energy; Future; Goals; Health; Human; Human Development; Inflammation; Integral Membrane Protein; Investigation; Label; Lead; Ligand Binding; Lipids; Liquid substance; Malignant Neoplasms; Measurement; Measures; Membrane; Methodology; Methods; Modeling; Molecular; Molecular Conformation; Mutate; Mutation; Nature; Pathogenicity; Pathology; Pathway interactions; Phosphotransferases; Play; Point Mutation; Positioning Attribute; Protein Family; Protocols documentation; Qi; Receptor Activation; Receptor Protein-Tyrosine Kinases; Research; Research Personnel; Resolution; Role; Signal Transduction; Signaling Protein; Structure; Techniques; Time; Transmembrane Domain; Tropomyosin; Validation; Vesicle; Visualization; base; computer studies; dimer; experimental study; gain of function; improved; insight; method development; mimetics; mutant; novel; open source; prevent; receptor; response; restraint; simulation; single molecule; solid state nuclear magnetic resonance; therapy development; user-friendly

Project Summary To sense the environment, cells rely on membrane-embedded receptors. The receptor tyrosine kinase (RTK) family of signaling proteins is large, diverse, and centrally important both to human development diseases and cancers. Evidence so far supports a model that signal passage through RTKs is initiated by a structural change in the extracellular domain and then conducted through the transmembrane (TMD) and juxtamembrane (JMD) domains to the cytoplasmic kinase domain. The receptors usually are activated in the dimer form. Numerous RTK mutations confer diseases, e.g. single point mutations in ~30% of residues of the TMD of the fibroblast growth factor receptor 3 (FGFR3) are pathogenic, while mutations of tropomyosin receptor kinase A can lead to cancers. Understanding the structural interactions of the FGFR3 and TrkA signaling TMD and JMD therefore is crucial for fundamental biology and for future development of therapies that may target these pathways. Atomistically resolved TMD+JMD dimer structures are the major objective of this project. Application of traditional computational and crystallographic methods is hindered by the fluid nature of the membrane environment. Our goal is to develop novel efficient computational methods that guide and maximally leverage NMR, FRET, and in-cell experimental data and apply these methods to capture the FGFR3 and TrkA TMD and TMD+JMD dimer structures for the wild type and mutated pathogenic forms. In Aim 1, we will combine our novel highly mobile membrane mimetic model, capable of spontaneously capturing candidate TMD dimer structures, with a novel minimally biased way of applying a reduced number of computational restraints based on experimental distance measurements. The resulting TMD dimer structures will be validated by comparing computed and experimentally measured parameters. These structures will reveal the role mutations play in RTK dynamics. In Aim 2, we will use our computational-experimental approach to determine the role that juxtamembrane domains play in RTK signaling. The resolved structures of the mutated dimers will facilitate understanding of the pathology and mechanisms of receptor activation. Our novel computational approaches combined with extended expertise of co-investigators and collaborators in NMR, FRET, RTK signaling, and membrane-associated phenomena, uniquely position us to develop and apply this methodology. We will also develop an open-source, user friendly workflow plugin for a widely-used software suite that will allow efficient use of the proposed protocols by the scientific community. Completion of the specific aims will increase our ability to efficiently gain structural information on RTKs and will open new research avenues for investigating mechanisms of transmembrane signaling in health and disease leading to development of new treatments.

项目概要 为了感知环境，细胞依靠膜嵌入的受体。受体酪氨酸激酶 (RTK) 信号蛋白家族庞大、多样，对人类发育疾病和 癌症。迄今为止的证据支持这样一个模型：通过 RTK 的信号通道是由结构变化引发的 在细胞外域，然后通过跨膜（TMD）和近膜（JMD）进行 域到细胞质激酶域。受体通常以二聚体形式被激活。很多的 RTK 突变会导致疾病，例如成纤维细胞 TMD 残基约 30% 发生单点突变 生长因子受体 3 (FGFR3) 具有致病性，而原肌球蛋白受体激酶 A 的突变可导致 到癌症。了解 FGFR3 和 TrkA 信号传导 TMD 和 JMD 的结构相互作用 对于基础生物学和针对这些途径的疗法的未来开发至关重要。 原子解析的TMD+JMD二聚体结构是该项目的主要目标。应用 传统的计算和晶体学方法受到膜的流体性质的阻碍 环境。我们的目标是开发新颖的高效计算方法来指导和最大限度地利用 NMR、FRET 和细胞内实验数据，并应用这些方法捕获 FGFR3 和 TrkA TMD 和 野生型和突变致病型的 TMD+JMD 二聚体结构。在目标 1 中，我们将结合我们的 新型高度移动的膜模拟模型，能够自发捕获候选TMD二聚体 结构，采用一种新颖的最小偏差方式来应用基于减少数量的计算限制 关于实验距离测量。由此产生的 TMD 二聚体结构将通过比较来验证 计算和实验测量的参数。这些结构将揭示突变在 RTK 动态。在目标 2 中，我们将使用计算实验方法来确定 近膜结构域在 RTK 信号传导中发挥作用。突变二聚体的解析结构将有助于 了解受体激活的病理学和机制。我们新颖的计算方法 结合共同研究人员和合作者在 NMR、FRET、RTK 信号传输和 膜相关现象，使我们能够开发和应用这种方法。我们还将 为广泛使用的软件套件开发一个开源、用户友好的工作流程插件，该插件将允许高效 科学界使用拟议的协议。完成具体目标将提高我们的 能够有效获取 RTK 的结构信息，并将为调查开辟新的研究途径 健康和疾病中的跨膜信号传导机制导致新疗法的开发。