Structure-guided optimization of light-activated chimeric G-protein-coupled receptors.

Autor: Tichy AM; 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., So WL; 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., Gerrard EJ; 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., Janovjak H; 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; Flinders Health and Medical Research Institute, College of Medicine and Public Health, Flinders University, 5042 Bedford Park/Adelaide, SA, Australia. Electronic address: harald.janovjak@flinders.edu.au.
Jazyk: angličtina
Zdroj: Structure (London, England : 1993) [Structure] 2022 Aug 04; Vol. 30 (8), pp. 1075-1087.e4. Date of Electronic Publication: 2022 May 18.
DOI: 10.1016/j.str.2022.04.012
Abstrakt: 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 β 2 AR 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.
Competing Interests: Declaration of interests The authors declare no competing interests.
(Copyright © 2022 Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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