Publication in eLife: a study by an EURIDOL team!

This page provides a sample of the publications of EURIDOL students and researchers of the Strasbourg Pain Initiative. Recent publications are presented first.

EURIDOL is pleased to announce the publication in the journal eLife of the article entitled “Affinity-guided labeling reveals P2X7 nanoscale membrane redistribution during BV2 microglial activation”, now available in its final version (version of record).

This work was carried out by the Ion Channel Engineering (ICI) team, which is part of the EURIDOL consortium, in collaboration with the Intelligent Nanoparticles (NPI) team, UMR 5297, and UPR CNRS 3212.

In this study, the researchers developed a novel chemical method to specifically biotinylate P2X7 receptors, enabling their labeling using streptavidin conjugated to Alexa Fluor 647. Applied to BV2 cells, a murine microglial cell line, and combined with super-resolution imaging techniques (dSTORM), this labeling strategy makes it possible to reveal and track the redistribution of P2X7 receptors at the single-molecule scale in response to inflammatory signaling.

The results demonstrate for the first time a clustering of P2X7 receptors following this inflammatory response. These findings open up new therapeutic perspectives, identifying the nanoscale reorganization of P2X7 receptors as a potential target for the treatment of inflammation.

 Like of the publication ! 

Affinity-guided labeling strategy for P2X7. (A) Schematic representation of the P2X7 labeling strategy using ligand-directed N-cyanomethyl NASA chemistry. A P2X7 ligand (Lg) binds to allosteric sites on the receptor, bringing the N-cyanomethyl NASA warhead into close proximity with endogenous lysine (K) residues. This spatial arrangement enables the covalent transfer of a biotin moiety via amide bond formation, resulting in highly specific biotinylation of P2X7. The biotin tag allows super-resolution imaging of nanoscale P2X7 localization using a Streptavidin-Alexa 647 probe (Strept-A 647). Notably, following biotin transfer, the ligand can dissociate from P2X7, leaving the allosteric sites unoccupied. (B) Crystal structure of panda P2X7 (pdP2X7) shown in ribbon representation, bound to AZ10606120 depicted as spheres (PDB:5U1W; Karasawa and Kawate, 2016). One of the three ATP-binding sites and the approximate location of the membrane are also indicated. Inset, enlarged view of the AZ10606120-binding pocket, rotated 180°. Distances (in Å) between the α-carbon of selected lysines and the hydroxyl group of AZ10606120 are displayed. Note that K300 is not visible in this view. (C) Chemical structure of X7-uP.