pH regulation of cell surface receptors

项目摘要 重合信号是蜂窝通信的基本特征。通过检测同时输入， 细胞可以在复杂的化学环境中从噪声中过滤信号，以产生适当的反应。在许多 在生物和病理背景下，一致的pH信号调节单个蛋白质的活性， 信令网络例子包括成熟内体中的酸信号，炎症区， 突触和肿瘤微环境。我们的长期目标是了解这些巧合 pH信号通过不同类型的细胞表面受体调节生物学。 我们已经制定了一个全面和雄心勃勃的研究计划，研究质子门控 （H+门控）符合检测G蛋白偶联受体（GPCR），最大和最 治疗靶向类的跨膜受体在人类。超过800种GPCR检测到 丰富多样的输入，包括H+。尽管一些pH敏感GPCR仅由质子激活， 我们已经表明H+门控符合检测是GPCR调节的更常见的特征。 在这种质子感测模式中，GPCR激动和/或抑制同时由pH调节。 我们努力阐明这种控制GPCR活性的背景依赖性机制， 建立了一个新的前沿细胞信号生物学，可能是相关的所有受体类别。 我们提出的研究计划的广泛目标是深入了解 H+门控信号和药理学用于各种GPCR。通过创建创新的湿实验室， 计算技术，开发尖端的细胞模型，并建立大型GPCR库 在基于细胞的检测系统中，我们的实验室可以广泛研究pH对GPCR信号传导的影响。因此，在本发明中， 我们可以使用我们的酵母为基础的分析雄心勃勃的GPCR和配体作为pH值的函数， DCyFIR平台和人类细胞模型。我们提出的研究计划包括三个项目 协同利用这些独特能力的领域：H+门控GPCR符合检测 代谢物和药物，通过GPCR对分泌肽和蛋白质传感的pH调节，以及pH- 智能纳米抗体研究工具和GPCR的治疗线索。 在接下来的五年里，我们的目标是阐明H+门控巧合的机制， 检测调节内源性和人工激动剂、抑制剂、调节剂的选择性， 药物和构象选择性纳米抗体探针，用于相当大一部分的人GPCRome。 我们预计这些努力将使我们能够设计和重新使用一系列治疗导线， 用于选择性靶向、控制和研究GPCR的探索性探针和药理学工具 在离散生理pH值下的信号传导机制。因此，我们期待我们雄心勃勃的研究 该计划将为酸中毒情况下的GPCR生物学和药理学建立新的范例。

PROJECT SUMMARY Coincident signals are an essential feature of cellular communication. By detecting simultaneous inputs, cells can filter signal from noise in complex chemical environments to mount proper responses. In many biological and pathological contexts, coincident pH signals regulate the activity of individual proteins and signaling networks. Examples include acidotic signals in maturing endosomes, inflammatory zones, synapses, and tumor microenvironments. Our long-term objective is to understand how these coincident pH signals regulate biology through different classes of cell surface receptors. We have developed a comprehensive and ambitious research program for studying proton-gated (H+-gated) coincidence detection by G protein-coupled receptors (GPCRs), the largest and most therapeutically targeted class of transmembrane receptors in humans. More than 800 GPCRs detect a rich diversity of inputs, including H+. Although a few pH-sensing GPCRs are activated by protons alone, we have shown that H+-gated coincidence detection is a far more common feature of GPCR regulation. In this mode of proton sensing, GPCR agonism and/or inhibition is concurrently modulated by pH. Our efforts to illuminate this context-dependent mechanism for controlling GPCR activity have led us to establish a new frontier in cell signaling biology that is likely relevant to all receptor classes. The broad objective of our proposed research program is to pursue an in-depth understanding of H+-gated signaling and pharmacology for a wide variety of GPCRs. By creating innovative wet-lab and computational technologies, developing cutting-edge cell models, and building large libraries of GPCRs in cell-based assay systems, our lab can extensively study the effects of pH on GPCR signaling. As such, we can profile ambitious numbers of GPCRs and ligands as a function of pH using our yeast based DCyFIR platform and human cell models. Our proposed program of research comprises three project areas that synergistically utilize these unique capabilities: H+-gated GPCR coincidence detection of metabolites and drugs, pH regulation of secreted peptide and protein sensing by GPCRs, and pH- intelligent nanobody research tools and therapeutic leads for GPCRs. Over the next five years, our goal is to illuminate the mechanisms by which H+-gated coincidence detection regulates the selectivity of endogenous and artificial agonists, inhibitors, modulators, approved drugs, and conformationally-selective nanobody probes for a sizeable fraction of the human GPCRome. We anticipate these efforts will enable us to both design and repurpose an array of therapeutic leads, exploratory probes, and pharmacological tools for selectively targeting, controlling, and studying GPCR signaling mechanisms at discrete physiologic pH values. As such, we anticipate our ambitious research program will establish a new paradigm for GPCR biology and pharmacology in acidotic scenarios.