Manipulation of ion channel gating with photoswitchable tweezers.

Fiche publication

Date publication

janvier 2021

Journal

Methods in enzymology

Auteurs

Membres identifiés du Cancéropôle Est :
Dr GRUTTER Thomas

Tous les auteurs :
Dunning K, Grutter T

Lien Pubmed

https://www.ncbi.nlm.nih.gov/pubmed/34099179

Résumé

The modulation of ion channel activity is of central importance within the nervous system, and an in-depth understanding of how such activity occurs on the molecular level is of prime importance for enhancing our understanding of neuronal systems in physiological and pathological states. The use of light as a stimulus has presented the unique opportunity to study these dynamic processes with exquisite spatiotemporal control. We have developed the photoswitchable tweezers method, an optogenetic pharmacology-based technique which relies on the use of a photoswitchable crosslinker as "tweezers" to manipulate the molecular movements involved in ion channel functionalities. Not only does this allow optical control of ion channel activity, but also investigation into the molecular motions and inter-residue distances implicated in such activity. In this chapter we discuss the principles behind the photoswitchable tweezers method, its strategic design and the key experimental steps involved in this technique, using purinergic P2X2 receptor as a case study system.