Australian Regenerative Medicine Institute (ARMI), Faculty of Medicine, Nursing and Health Sciences, Monash University, 3800 Clayton/Melbourne, VIC, Australia; European Molecular Biology Laboratory Australia (EMBL Australia), Monash University, 3800 Clayton/Melbourne, VIC, Australia.
Australian Regenerative Medicine Institute (ARMI), Faculty of Medicine, Nursing and Health Sciences, Monash University, 3800 Clayton/Melbourne, VIC, Australia; European Molecular Biology Laboratory Australia (EMBL Australia), Monash University, 3800 Clayton/Melbourne, VIC, Australia; Commonwealth Scientific and Industrial Research Organisation, Synthetic Biology Future Science Platform, Monash University, 3800 Clayton/Melbourne, VIC, Australia.
Structure. 2022 Aug 4;30(8):1075-1087.e4. doi: 10.1016/j.str.2022.04.012. Epub 2022 May 18.
G-protein-coupled receptors (GPCRs) are the largest human receptor family and involved in virtually every physiological process. One hallmark of their function is specific coupling to selected signaling pathways. The ability to tune this coupling would make development of receptors with new capabilities possible. Complexes of GPCRs and G-proteins have recently been resolved at high resolution, but this information was in only few cases harnessed for rational receptor engineering. Here, we demonstrate structure-guided optimization of light-activated OptoXRs. Our hypothesis was that incorporation of GPCR-Gα contacts would lead to improved coupling. We first evaluated structure-based alignments for chimeric receptor fusion. We then show in a light-activated βAR that including Gα contacts increased signaling 7- to 20-fold compared with other designs. In turn, contact elimination diminished function. Finally, this platform allowed optimization of a further OptoXR and spectral tuning. Our work exemplifies structure-based OptoXR development for targeted cell and network manipulation.
G 蛋白偶联受体（GPCRs）是人类最大的受体家族，参与几乎所有的生理过程。其功能的一个显著特点是与特定的信号通路特异性偶联。调节这种偶联的能力将使具有新功能的受体的开发成为可能。最近，高分辨率解析了 GPCR 和 G 蛋白复合物，但在少数情况下，这些信息被用于合理的受体工程。在这里，我们展示了基于结构的光激活 OptoXR 的优化。我们的假设是，掺入 GPCR-Gα 接触会导致更好的偶联。我们首先评估了嵌合受体融合的基于结构的比对。然后，我们在光激活的βAR 中表明，与其他设计相比，包括 Gα 接触可将信号增加 7 到 20 倍。反过来，接触消除会降低功能。最后，该平台允许进一步优化 OptoXR 并进行光谱调整。我们的工作为例说明了基于结构的 OptoXR 开发，用于靶向细胞和网络操作。
