The role of dynamics in GPCR and arrestin allostery

动力学在 GPCR 和抑制蛋白变构中的作用

• 批准号: 10276858
• 负责人: Joshua James Ziarek
• 金额: $ 39.63万
• 依托单位: TRUSTEES OF INDIANA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-15 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10276858
• 关键词: Agonist; Architecture; Arrestins; Binding; Cardiovascular Diseases; Complement; Complex; Cryoelectron Microscopy; Crystallization; Entropy; FDA approved; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; GTP-Binding Proteins; Gastrointestinal Diseases; Goals; Inflammatory; Label; Ligand Binding; Ligands; Lung diseases; Malignant Neoplasms; Measurement; Measures; Membrane Proteins; Molecular; Molecular Conformation; Motion; NMR Spectroscopy; Peptides; Pharmaceutical Preparations; Protein Family; Proteins; Proxy; Receptor Signaling; Relaxation; Research; Research Project Grants; Resolution; Roentgen Rays; Role; Signal Transduction; Signaling Molecule; Structural Models; Structure; Techniques; Therapeutic; Translating; conformational conversion; drug development; extracellular; human disease; mutant; novel therapeutics; programs; receptor; response; virtual

PROJECT SUMMARY/ABSTRACT GPCRs form the largest membrane protein family and also dominate the therapeutic market as targets for more than 30% of FDA-approved drugs. These drugs act on a broad spectrum of indications from cancer to inflammatory, cardiovascular, respiratory and gastrointestinal disease. GPCRs are archetypical allosteric proteins that translate extracellular ligand stimulation into an intracellular response. In contrast to the binary “on or off” response of most signaling molecules, GPCRs possess a ligand-independent basal activity that can be increased or decreased upon ligand binding, and then further regulated by allosteric modulators. Activated receptors transduce signals through G protein and arrestin proteins equally (balanced signaling) or selectively (biased signaling). Taken together, a single receptor may specifically recognize several ligands and respond uniquely to each, creating a complex conformational landscape. The last decade has seen nearly 300 X-ray and cryo-EM structures have greatly expanded our view of GPCR architecture and function; however, the molecular mechanisms of basal activity, partial agonism, biased signaling, and allosteric modulation can only be partially derived from these structures. The principles of class Monod-Wyman-Changeux (MWC) and dynamically-drive (DD) allostery can significantly enhance our understanding of, and ability to tune, GPCR signaling. Whereas classic MWC conformational allostery is relatively simple to infer from X-ray or cryo-EM structural models, DD allostery is far more difficult to measure experimentally. Advances in NMR spectroscopy relaxation measurements have empirically-demonstrated that sidechain methyl dynamics can be used as a proxy for conformational entropy (i.e. DD allostery). In addition, it remains the only technique capable of quantifying atomic-resolution motions across the picosecond to second timescale and in many cases can detect states populated as little as 0.5%. We propose two research projects aimed at exploring the allosteric role of MWC and DD allosteric mechanisms in the activation of 1) peptide-binding GPCRs and 2) arrestin. There is immense therapeutic potential in the ability to tune receptor signaling using partial or biased agonists in contrast to full agonists/antagonists – and it remains virtually untapped. Our proposal provides a much- needed complement to decades of functional mutant screens, EPR, fluorescent labels, and high-resolution structures. In the long-term, the goal of our research program is to describe the conformational transitions and dynamics of the greater GPCR superfamily from the inactive state to the active state unhindered by crystal contacts or stabilizing proteins.

项目摘要/摘要 GPCR构成了最大的膜蛋白家族，并在治疗市场中占据主导地位 超过30％的FDA批准药物。这些药物以从癌症到癌症的广泛适应症作用 GPCR是原型变构的 将细胞外配体刺激转化为细胞内反应的蛋白质。与二进制 或OFF”大多数信号分子的响应，GPCR具有与配体无关的基本活性，可以是 配体结合后增加或减少，然后由变构调节剂进一步调节。活性 接收器通过G蛋白和阻止蛋白（平衡信号传导）或选择性地传递信号 （偏置信号）。综上 独特地对每个人创造了复杂的构象景观。最近十年有近300 X射线 冷冻EM结构极大地扩展了我们对GPCR架构和功能的看法。但是， 基本活性，部分激动剂，偏置信号传导和变构调制的分子机制只能 部分源自这些结构。班级单人卫生 - 更改（MWC）的原则和 动态驱动（DD）变构可以显着增强我们对GPCR的理解和调整能力 信号。而经典的MWC构象变构相对简单，从X射线或冰冻EM推断 结构模型，DD变构很难通过实验测量。 NMR光谱的进步 放松测量已在经验上证明了侧chain甲基动力学可以用作 代理构象熵（即DD变构）。此外，它仍然是唯一能够 量化Picsecond至第二时间尺度的原子分辨率运动，在许多情况下可以 检测人口少的0.5％。我们提出了两个旨在探索变构的研究项目 MWC和DD变构机制在1）肽结合GPCR和2）阻止素中的作用。 使用部分或偏见的激动剂调节受体信号传导的能力具有巨大的治疗潜力 与完整的激动剂/拮抗剂相反，它几乎尚未开发。我们的建议提供了很多 需要数十年的功能突变屏幕，EPR，荧光标签和高分辨率的补充 结构。从长远来看，我们的研究计划的目标是描述构象转变和 从无效状态到活跃状态的大型GPCR超家族的动力学不受晶体的影响 接触或稳定蛋白质。