Identification of signal transduction pathways used by orphan G protein-coupled receptors
Identification of signal transduction pathways used by orphan G protein-coupled receptors
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DOI：
10.1089/15406580360545053
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发表时间：
2003-04-01
影响因子：
1.8
通讯作者：
McAllister, G
中科院分区：
文献类型：
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作者：
Bresnick, JN;Skynner, HA;McAllister, G
GPCR 超家族具有多种生物学作用，可转导来自一系列刺激的信号，从视蛋白的光子识别到神经元功能的神经递质调节。在许多已识别的编码 GPCR 的基因中，超过 130 个是孤儿受体（即它们的内源配体未知），并且该子集代表了多种疾病的药物干预的假定新治疗靶点。作为药物发现的第一步，确定这些新鉴定的受体的生物学功能至关重要，因此天然配体的鉴定是首要目标。有几种已建立的方法可以做到这一点，但许多方法都有缺点，并且通常需要对受体如何发挥作用有深入的了解。这里描述的技术在活细胞中利用基于转录的报告基因测定。这允许确定任何给定 oGPCR 使用的信号转导途径，而无需事先了解内源配体。因此，这可以减少以多种形式（即 Galpha(s)、Galpha(i/0) 和 Galpha(q) 测定）筛选给定受体的配体所涉及的冗余工作，并确保目标受体在适当激活的情况下能够发出信号。这些知识通常需要费力才能获得，并且对于几乎所有的 oGPCR 来说，此类信息尚不可用。该技术还可用于开发 oGPCR 的反向激动剂以及激动剂敏感的高通量检测。使用许多已知的 GPCR 证明了该方法的准确性。显示了 GPR3、GPR12、GPR19、GPR21 和 HG55 oGPCR 的可能信号传导途径，并开发了 GPR26 受体的高通量测定。这里概述的用于阐明 oGPCR 信号转导途径和开发功能测定的方法应该会加快这一潜在治疗有用受体组的配体鉴定过程。
The superfamily of GPCRs have diverse biological roles, transducing signals from a range of stimuli, from photon recognition by opsins to neurotransmitter regulation of neuronal function. Of the many identified genes encoding GPCRs, >130 are orphan receptors (i.e., their endogenous ligands are unknown), and this subset represents putative novel therapeutic targets for pharmaceutical intervention in a variety of diseases. As an initial step toward drug discovery, determining a biological function for these newly identified receptors is of vital importance, and thus identification of a natural ligand(s) is a primary aim. There are several established methods for doing this, but many have drawbacks and usually require some in-depth knowledge about how the receptor functions. The technique described here utilizes a transcription-based reporter assay in live cells. This allows the determination of the signal transduction pathway any given oGPCR uses, without any prior knowledge of the endogenous ligand. This can therefore reduce the redundancy of effort involved in screening ligands at a given receptor in multiple formats (i.e., Galpha(s), Galpha(i/0), and Galpha(q) assays), as well as ensuring that the receptor targeted is capable of signaling if appropriately activated. Such knowledge is often laboriously obtained, and for almost all oGPCRs, this kind of information is not yet available. This technology can also be used to develop inverse agonist as well as agonist sensitive high throughput assays for oGPCRs. The veracity of this approach is demonstrated, using a number of known GPCRs. The likely signaling pathways of the GPR3, GPR12, GPR19, GPR21, and HG55 oGPCRs are shown, and a high throughput assay for GPR26 receptors developed. The methods outlined here for elucidation of the signal transduction pathways for oGPCRs and development of functional assays should speed up the process of identification of ligands for this potentially therapeutically useful group of receptors.