Comparison of Agonist Activity between CB1 and CB2 Receptors with Orthosteric Site Mutations.

Receptors (Basel, Switzerland) Pub Date : 2024-09-01 Epub Date: 2024-08-06 DOI:10.3390/receptors3030018
Christina A Brust, Matthew A Swanson, Christos Iliopoulos Tsoutsouvas, Snezana T Dimova, Vuong Q Dang, Edward L Stahl, Jo-Hao Ho, Spyros P Nikas, Alexandros Makriyannis, Laura M Bohn
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Abstract

Human endocannabinoid signaling is primarily mediated by the cannabinoid receptors, CB1 and CB2, which are G protein-coupled receptors (GPCRs). These receptors have been linked to a variety of physiological processes and are being pursued as prospective drug targets due to their potential in treating pain and inflammation. However, because of their homology and shared signaling mechanisms, investigating the individual physiological roles of these receptors and designing subtype-selective ligands has been challenging. Using active-state CB1 and CB2 structures as guides, homologous residues within the orthosteric pocket of each receptor were mutated to alanine to test whether they equally impair CB1 and CB2 activity in response to two high-affinity, nonselective agonists (CP55,940 and AM12033). Interestingly, mutating the Y5.39 position impairs CB1 but not CB2 function. Conversely, mutating residue C6.47 improves CB1 but impairs CB2 signaling. TheF7.35A mutation leads to a decrease in CP55,940 potency at CB1 and impairs internalization; however, AM12033 gains potency and promotes CB1 internalization. In CB2, mutation of F7.35A decreases the potency of CP55,940 and neither agonist induces internalization. These observations provide some insight into functional sensitivity of CB1 and CB2 to different agonists when conserved residues are mutated in the orthosteric pocket.

CB1和CB2受体与正位位点突变的激动剂活性比较。
人内源性大麻素信号主要由大麻素受体CB1和CB2介导,它们是G蛋白偶联受体(gpcr)。这些受体与多种生理过程有关,由于它们在治疗疼痛和炎症方面的潜力,正被用作潜在的药物靶点。然而,由于它们的同源性和共享的信号机制,研究这些受体的个体生理作用和设计亚型选择性配体一直具有挑战性。以活性状态CB1和CB2结构为指导,在每个受体的正位袋内的同源残基突变为丙氨酸,以测试它们是否在响应两种高亲和力,非选择性激动剂(CP55,940和AM12033)时同样损害CB1和CB2的活性。有趣的是,突变Y5.39位点会损害CB1,但不会损害CB2的功能。相反,突变残基C6.47改善了CB1,但损害了CB2信号传导。f7.35 a突变导致CP55,940在CB1的效力降低,并损害内化;然而,AM12033获得效力并促进CB1内化。在CB2中,F7.35A的突变降低了CP55,940的效力,两种激动剂都不能诱导内化。这些观察结果为CB1和CB2对不同激动剂的功能敏感性提供了一些见解,当保守残基在矫形袋中发生突变时。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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