SNAP-Tagged Nanobodies Enable Reversible Optical Control of a G Protein-Coupled Receptor via a Remotely Tethered Photoswitchable Ligand

ACS Chemical Biology
Helen FarrantsJohannes Broichhagen

Abstract

G protein-coupled receptors (GPCRs) mediate the transduction of extracellular signals into complex intracellular responses. Despite their ubiquitous roles in physiological processes and as drug targets for a wide range of disorders, the precise mechanisms of GPCR function at the molecular, cellular, and systems levels remain partially understood. To dissect the function of individual receptor subtypes with high spatiotemporal precision, various optogenetic and photopharmacological approaches have been reported that use the power of light for receptor activation and deactivation. Here, we introduce a novel and, to date, most remote way of applying photoswitchable orthogonally remotely tethered ligands by using a SNAP-tag fused nanobody. Our nanobody-photoswitch conjugates can be used to target a green fluorescent protein-fused metabotropic glutamate receptor by either gene-free application of purified complexes or coexpression of genetically encoded nanobodies to yield robust, reversible control of agonist binding and subsequent downstream activation. By harboring and combining the selectivity and flexibility of both nanobodies and self-labeling proteins (or suicide enzymes), we set the stage for targeting endogenous receptors i...Continue Reading

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Citations

Jul 11, 2019·Nature Reviews. Neuroscience·Pierre PaolettiAlexandre Mourot
Aug 6, 2020·Proceedings of the National Academy of Sciences of the United States of America·Conor McMahonRobert J Lefkowitz
May 1, 2020·Nature Communications·Ross W ChelohaHidde L Ploegh
Sep 2, 2020·The Journal of Biological Chemistry·Ross W ChelohaHidde L Ploegh
Dec 17, 2020·Biomolecules·Eline SoetensXavier Saelens
Mar 19, 2021·Cell Chemical Biology·Vanessa A GutzeitJohannes Broichhagen
Feb 12, 2021·Chembiochem : a European Journal of Chemical Biology·Johannes Broichhagen, Nicole Kilian
Aug 12, 2019·Trends in Pharmacological Sciences·Sarah MondoloniAlexandre Mourot

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