Adhesion G protein-coupled receptor (aGPCR) signaling influences development and homeostasis in a wide range of tissues. In the current model for aGPCR signaling, ligand binding liberates a conserved sequence that acts as an intramolecular, tethered agonist (TA), yet this model has not been evaluated systematically for all aGPCRs. Here, we assessed the TA-dependent activities of all 33 aGPCRs in a suite of transcriptional reporter, G protein activation, and β-arrestin recruitment assays using a new fusion protein platform. Strikingly, only ∼50% of aGPCRs exhibited robust TA-dependent activation, and unlike other GPCR families, aGPCRs showed a notable preference for G_12/13 signaling. AlphaFold2 predictions assessing TA engagement in the predicted intramolecular binding pocket aligned with the TA dependence of the cellular responses. This dataset provides a comprehensive resource to inform the investigation of all human aGPCRs and for targeting aGPCRs therapeutically.

粘附 G 蛋白偶联受体 (aGPCR) 信号传导影响多种组织的发育和稳态。在当前的 aGPCR 信号传导模型中，配体结合释放出一个保守序列，充当分子内束缚激动剂 (TA)，但该模型尚未针对所有 aGPCR 进行系统评估。在这里，我们使用新的融合蛋白平台在一套转录报告基因、G 蛋白激活和 β-arrestin 募集测定中评估了所有 33 个 aGPCR 的 TA 依赖性活性。引人注目的是，只有约 50% 的 aGPCR 表现出强大的 TA 依赖性激活，并且与其他 GPCR 家族不同，aGPCR 对 G _12/13信号传导表现出显着的偏好。 AlphaFold2 预测评估 TA 在预测的分子内结合口袋中的参与情况，与细胞反应的 TA 依赖性相一致。该数据集提供了全面的资源，可为所有人类 aGPCR 的研究以及针对 aGPCR 的治疗提供信息。