Zheng Yuan, Jiang Dan, Lu Yan, Zhang Chao, Huang Shen-Ming, Lin Haocheng, Zhang Daolai, Guo Shengchao, Han Jifei, Chen Jun, He Yaxuan, Zhang Mingxiang, Gao Yanhui, Guo Yongyuan, Wei Ran, Xia Ming, Qin Yingying, Liu Zhaoqian, Yang Fan, Ge Shaohua, Yi Fan, Yu Xiao, Lin Hui, Xiao Peng, Sun Jin-Peng, Feng Shiqing
Department of Physiology and Pathophysiology, School of Basic Medical Sciences, State Key Laboratory of Vascular Homeostasis and Remodeling, Peking University, Beijing, China.
Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China.
Nat Chem Biol. 2025 May 15. doi: 10.1038/s41589-025-01896-2.
Allosteric modulation of receptor responses to endogenous agonists has therapeutic value, maintaining ligand profiles, reducing side effects and restoring mutant responses. Adhesion G-protein-coupled receptors (aGPCRs), with large N termini, are ideal for allosteric modulator development. We designed a nanobody strategy targeting ADGRG2 N-terminal fragments and got a specific nanobody Nb23-bi, which promoted dehydroepiandrosterone (DHEA)-induced ADGRG2 activation and reversed mutant-induced dysfunctions. By combining structural characterization, crosslinking mass spectrometry, mutational analysis and molecular dynamics simulations, we clarified the allosteric mechanism of how the Nb23-bi modulates conformational changes in the DHEA-binding pocket. Animal studies showed that Nb23-bi promoted the response of DHEA in alleviating testicular inflammation and reversing mutant defects. In summary, we developed an allosteric nanobody of ADGRG2 and gained insights into its functions in reversing disease-associated dysfunctions. Our study may serve as a template for developing allosteric modulators of other aGPCRs for biological and therapeutic purposes.
受体对内源性激动剂反应的变构调节具有治疗价值，可维持配体特征、减少副作用并恢复突变体反应。具有大的N端的粘附G蛋白偶联受体（aGPCRs）是变构调节剂开发的理想选择。我们设计了一种靶向ADGRG2 N端片段的纳米抗体策略，并获得了一种特异性纳米抗体Nb23-bi，它促进了脱氢表雄酮（DHEA）诱导的ADGRG2激活，并逆转了突变体诱导的功能障碍。通过结合结构表征、交联质谱、突变分析和分子动力学模拟，我们阐明了Nb23-bi调节DHEA结合口袋构象变化的变构机制。动物研究表明，Nb23-bi促进了DHEA在减轻睾丸炎症和逆转突变体缺陷方面的反应。总之，我们开发了一种ADGRG2的变构纳米抗体，并深入了解了其在逆转疾病相关功能障碍中的作用。我们的研究可为开发用于生物学和治疗目的的其他aGPCRs变构调节剂提供模板。
