Screening β-Arrestin Recruitment for the Identification of Natural Ligands for Orphan G-Protein-Coupled Receptors
Screening β-Arrestin Recruitment for the Identification of Natural Ligands for Orphan G-Protein-Coupled Receptors
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DOI：
10.1177/1087057113475480
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发表时间：
2013-06-01
影响因子：
--
通讯作者：
Rees, Steve
中科院分区：
文献类型：
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作者：
Southern, Craig;Cook, Jennifer M.;Rees, Steve
多种 G 蛋白偶联受体 (GPCR) 筛选技术已成功地将许多 GPCR 与其同源配体结合起来。 GPCR 介导的 β-arrestin 募集现已被认为是一种独特的细胞内信号传导途径，并且配体-受体相互作用可能显示出相对于经典 GPCR 信号传导途径偏向 β-arrestin。我们假设未能识别剩余孤儿 GPCR 的天然配体可能是 β-arrestin 信号传导偏向的结果。为了研究这一点，我们组装了 10500 个候选配体，并使用 PathHunter beta-arrestin 招募技术筛选了 82 个 GPCR。通过包含配体受体以及检测和确认这些已建立的配体-受体配对来验证高质量的筛选测定。我们描述了候选内源孤儿 GPCR 配体和许多新型替代配体。然而，对于大多数研究的孤儿受体，β-抑制蛋白募集的测量并没有导致从我们的筛选组中鉴定出同源配体。 beta-Arrestin 募集代表了一种强大的 GPCR 筛选技术，而配体偏向信号传导正在成为 GPCR 生物学的一种治疗上可利用的特征。孤儿 GPCR 的同源配体的鉴定以及受体可能存在的程度，以优先通过 β-arrestin 响应其天然配体发出信号，仍有待确定。
A variety of G-protein-coupled receptor (GPCR) screening technologies have successfully partnered a number of GPCRs with their cognate ligands. GPCR-mediated beta-arrestin recruitment is now recognized as a distinct intracellular signaling pathway, and ligand-receptor interactions may show a bias toward beta-arrestin over classical GPCR signaling pathways. We hypothesized that the failure to identify native ligands for the remaining orphan GPCRs may be a consequence of biased beta-arrestin signaling. To investigate this, we assembled 10 500 candidate ligands and screened 82 GPCRs using PathHunter beta-arrestin recruitment technology. High-quality screening assays were validated by the inclusion of liganded receptors and the detection and confirmation of these established ligand-receptor pairings. We describe a candidate endogenous orphan GPCR ligand and a number of novel surrogate ligands. However, for the majority of orphan receptors studied, measurement of beta-arrestin recruitment did not lead to the identification of cognate ligands from our screening sets. beta-Arrestin recruitment represents a robust GPCR screening technology, and ligand-biased signaling is emerging as a therapeutically exploitable feature of GPCR biology. The identification of cognate ligands for the orphan GPCRs and the extent to which receptors may exist to preferentially signal through beta-arrestin in response to their native ligand remain to be determined.