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: 10582241
• 负责人: Taras V. Pogorelov
• 金额: $ 11.32万
• 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/10582241
• 关键词: Award; Biology; Cells; Communities; Computer software; Computing Methodologies; Data; Development; Diabetes Mellitus; Disease; Environment; Extracellular Domain; FGFR3 gene; Family; Fluorescence Resonance Energy Transfer; Funding; Future; Goals; Health; Human Development; Inflammation; Lead; Liquid substance; Malignant Neoplasms; Measurement; Measures; Membrane; Methodology; Methods; Modeling; Mutate; Mutation; Nature; Pathogenicity; Pathology; Pathway interactions; Phosphotransferases; Play; Point Mutation; Positioning Attribute; Protocols documentation; Receptor Activation; Receptor Protein-Tyrosine Kinases; Research; Research Personnel; Role; Signal Transduction; Signaling Protein; Structure; Tropomyosin; base; dimer; improved; mimetics; novel; open source; receptor; restraint; therapy development; user-friendly

Project Summary of the Funded Award (R01GM141298) 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.

资助奖的项目摘要（R01GM141298） 为了感知环境，细胞依赖于膜上的受体。受体酪氨酸 信号蛋白的激酶（RTK）家族对人的大型，多样化和中心很重要 发育疾病和癌症。到目前为止的证据支持一个信号通道的模型 通过RTK是由细胞外域的结构变化引发的，然后进行 通过跨膜（TMD）和juxtammbrane（JMD）结构域到达细胞质激酶 领域。通常以二聚体形式激活受体。许多RTK突变会议 疾病，例如成纤维细胞生长的TMD残基约30％的单点突变 因子受体3（FGFR3）是致病性的，而tropomyosin受体激酶的突变可以 导致癌症。了解FGFR3和TRKA信号TMD的结构相互作用 因此，JMD对于基本生物学和未来的疗法发展至关重要 可能针对这些途径。原子上解决的TMD+JMD二聚体结构是主要的 这个项目的目标。传统计算和晶体学方法的应用是 受膜环境的流体性质阻碍。我们的目标是发展新颖的效率 指导和最大利用NMR，FRET和细胞内实验的计算方法 数据并应用这些方法来捕获FGFR3和TRKA TMD和TMD+JMD二聚体 野生型和突变的致病形式的结构。在AIM 1中，我们将结合小说 高移动的膜模拟模型，能够自发捕获候选TMD 二聚体结构，采用新颖的偏差方式来应用减少数量 基于实验距离测量值的计算约束。由此产生的TMD 二聚体结构将通过比较计算和实验测量来验证 参数。这些结构将揭示突变在RTK动力学中的作用。在AIM 2中，我们 将使用我们的计算实验方法来确定置术的作用 域在RTK信号中播放。突变二聚体的解决结构将有助于 了解受体激活的病理和机制。我们的新颖计算 方法结合了NMR的共同投资者和合作者的扩展专业知识， FRET，RTK信号传导和膜相关现象，独特地定位了我们发展 并应用此方法。我们还将开发一个开源的，用户友好的工作流插件 对于广泛使用的软件套件，该套件将有效地使用该协议 科学界。完成特定目标的完成将提高我们有效获得的能力 有关RTK的结构信息，并将为研究机制开放新的研究途径 跨膜信号在健康和疾病中导致新疗法的发展。