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: 10491306
• 负责人: Taras V. Pogorelov
• 金额: $ 30.85万
• 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/10491306
• 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.

项目摘要 为了感知环境，细胞依赖于膜上的受体。受体酪氨酸激酶（RTK） 信号蛋白的家族对人类发育疾病和 癌症。到目前为止的证据支持一个模型，即通过结构性变化启动了通过RTK的信号传递 在细胞外结构域中，然后通过跨膜（TMD）和近膜（JMD）进行 细胞质激酶结构域的结构域。通常以二聚体形式激活受体。很多的 RTK突变赋予疾病，例如〜30％的成纤维细胞TMD残基的单点突变 生长因子受体3（FGFR3）是致病性的，而肌球蛋白受体激酶A的突变可以铅 癌症。因此，了解FGFR3和TRKA信号TMD和JMD的结构相互作用 对于基本生物学以及可能针对这些途径的疗法的发展至关重要。 原子化解决的TMD+JMD二聚体结构是该项目的主要目标。应用 传统的计算方法和晶体学方法受膜的流体性质阻碍 环境。我们的目标是开发新颖的有效计算方法，以指导和最大程度地利用 NMR，FRET和细胞内实验数据，并应用这些方法来捕获FGFR3和TRKA TMD和 TMD+JMD二聚体结构，用于野生型和突变的致病形式。在AIM 1中，我们将结合我们的 新型高移动膜模拟模型，能够自发捕获候选TMD二聚体 结构，采用一种新颖的偏见方式，用于应用减少的基于计算的基础数量 实验距离测量。通过比较，将通过比较 计算和实验测量的参数。这些结构将揭示突变在 RTK动力学。在AIM 2中，我们将使用我们的计算实验方法来确定 Juxtammbrane域在RTK信号传导中播放。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NMR，FRET，RTK信号和合作者的扩展专业知识，并 膜相关现象，独特地定位了我们开发和应用这种方法。我们也会 为广泛使用的软件套件开发开源的，用户友好的工作流插件，该插件将允许有效 使用科学界提出的协议。具体目标的完成将增加我们的 能够有效获取RTK的结构信息，并将为研究的新研究途径开放 跨膜信号在健康和疾病中的机制，导致新治疗的发展。