pH regulation of cell surface receptors

项目总结 重合信号是蜂窝通信的基本特征。通过检测同时输入， 在复杂的化学环境中，细胞可以过滤噪音中的信号，从而做出适当的反应。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，并 信令网络。例如，成熟的内小体、炎症区、 突触和肿瘤微环境。我们的长期目标是了解它们是如何重合的 PH信号通过不同类别的细胞表面受体调节生物。 我们已经制定了一个全面而雄心勃勃的研究计划来研究质子门 最大和最多的G蛋白偶联受体(GPCRs)的(H+门控)符合检测 治疗靶向的人类跨膜受体类。超过800个GPCR检测到 丰富多样的投入，包括H+。尽管少数pH敏感的GPCR仅由质子激活， 我们已经证明，H+门控符合检测是GPCR调节的一个更常见的特征。 在这种质子传感模式中，GPCR的兴奋和/或抑制同时受到pH的调节。 我们努力阐明这一依赖于上下文的控制GPCR活动的机制，导致我们 在可能与所有受体类别相关的细胞信号生物学领域建立一个新的前沿。 我们建议的研究计划的广泛目标是深入了解 多种GPCRs的H+门控信号和药理学。通过创建创新的湿实验室和 计算技术，开发尖端细胞模型，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地研究pH对GPCR信号的影响。因此， 我们可以使用我们的酵母基，将庞大的GPCRs和配体数量描述为pH的函数 DCyFIR平台和人体细胞模型。我们建议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，GPCRs对分泌肽和蛋白质的pH调节，以及pH- 用于GPCRs的智能纳米体研究工具和治疗线索。 在接下来的五年里，我们的目标是阐明H+门控符合的机制 检测调节批准的内源性和人工激动剂、抑制剂、调节剂的选择性 药物，以及构象选择性纳米体探针对相当大一部分人的GPC罗马。 我们预计这些努力将使我们能够设计和重新定位一系列治疗线索， 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 在离散的生理pH值下的信号机制。因此，我们期待着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一个新的范例。

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.