CHAPPE, Yann; PIERREDON, Sandra; JOUSHOMME, Alexandre; MOLLE, Pablo; GARENNE, André; CANOVI, Anne; BARBEAU, Solene; POULLETIER DE GANNES, Florence; HURTIER, Annabelle; LAGROYE, Isabelle; DUCRET, Thomas; QUIGNARD, Jean-François; COMPAN, Vincent; PERCHERANCIER, Yann

doi:10.1073/pnas.2205207119

Laboratoire de l'intégration, du matériau au système [IMS]

PIERREDON, Sandra

JOUSHOMME, Alexandre

Laboratoire de l'intégration, du matériau au système [IMS]

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Article de revue

Ce document a été publié dans

Proceedings of the National Academy of Sciences of the United States of America. 2022-11-07, vol. 119, n° 46, p. e2205207119

Résumé en anglais

Whether ion channels experience ligand-dependent dynamic ion selectivity remains of critical importance since this could support ion channel functional bias. Tracking selective ion permeability through ion channels, however, remains challenging even with patch-clamp electrophysiology. In this study, we have developed highly sensitive bioluminescence resonance energy transfer (BRET) probes providing dynamic measurements of Ca2+ and K+ concentrations and ionic strength in the nanoenvironment of Transient Receptor Potential Vanilloid-1 Channel (TRPV1) and P2X channel pores in real time and in live cells during drug challenges. Our results indicate that AMG517, BCTC, and AMG21629, three well-known TRPV1 inhibitors, more potently inhibit the capsaicin (CAPS)-induced Ca2+ influx than the CAPS-induced K+ efflux through TRPV1. Even more strikingly, we found that AMG517, when injected alone, is a partial agonist of the K+ efflux through TRPV1 and triggers TRPV1-dependent cell membrane hyperpolarization. In a further effort to exemplify ligand bias in other families of cationic channels, using the same BRET-based strategy, we also detected concentration- and time-dependent ligand biases in P2X7 and P2X5 cationic selectivity when activated by benzoyl-adenosine triphosphate (Bz-ATP). These custom-engineered BRET-based probes now open up avenues for adding value to ion-channel drug discovery platforms by taking ligand bias into account.< Réduire

Mots clés en anglais

Transient Receptor Potential Channels

TRPV Cation Channels

Ligands

Capsaicin

Energy Transfer

Bias

URI

https://oskar-bordeaux.fr/handle/20.500.12278/170333

DOI

http://dx.doi.org/10.1073/pnas.2205207119

Project ANR

Dépasser les limites des méthodes existantes de criblage à haut débit et de caractérisation des canaux ioniques grâce au transfert d'énergie en résonance de bioluminescence - ANR-19-CE44-0010

Unités de recherche

IMS : Laboratoire de l'Intégration du Matériau au Système - UMR 5218

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Genetically-encoded BRET probes shed light on ligand bias–induced variable ion selectivity in TRPV1 and P2X5/7