Patch-clamp Coordinated Spectroscopy Shows P2X2 Receptor Permeability Dynamics Require Cytosolic Domain Rearrangements but Not Panx-1 Channels

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2008 Aug 12

PMID 18689682

Citations 49

Authors

Severine Chaumont

Baljit S Khakh

Affiliations

Soon will be listed here.

Abstract

ATP-gated P2X receptors display ion permeability increases within seconds of receptor activation as the channels enter the I(2) state, which is permeable to organic cations and dye molecules. The mechanisms underlying this important behavior are not completely understood. In one model, the I(2) state is thought to be due to opening of Pannexin-1 (Panx-1) channels, and, in the second, it is thought to be an intrinsic P2X property. We tested both models by measuring ion and dye permeability and used a patch-clamp coordinated spectroscopy approach to measure conformational changes. Our data show that Panx-1 channels make no detectable contribution to the P2X(2) receptor I(2) state. However, P2X(2) receptors display permeability dynamics, which are correlated with conformational changes in the cytosolic domain remote from the selectivity filter itself. Finally, the data illustrate the utility of a new approach, using tetracysteine tags and biarsenical fluorophores to measure site-specific conformational changes in membrane proteins.

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