Photo-switchable tweezers illuminate pore-opening motions of an ATP-gated P2X ion channel

• Main Authors: Chloé Habermacher, Adeline Martz, Nicolas Calimet, Damien Lemoine, Laurie Peverini, Alexandre Specht, Marco Cecchini, Thomas Grutter
• Format: Article
• Language: English
• Published: eLife Sciences Publications Ltd 2016-01-01
• Series: eLife
• Subjects: P2X receptors; azobenzene photoswitch; gating mechanism
• Online Access: https://elifesciences.org/articles/11050
• ISSN: 2050-084X

P2X receptors function by opening a transmembrane pore in response to extracellular ATP. Recent crystal structures solved in apo and ATP-bound states revealed molecular motions of the extracellular domain following agonist binding. However, the mechanism of pore opening still remains controversial. Here we use photo-switchable cross-linkers as ‘molecular tweezers’ to monitor a series of inter-residue distances in the transmembrane domain of the P2X2 receptor during activation. These experimentally based structural constraints combined with computational studies provide high-resolution models of the channel in the open and closed states. We show that the extent of the outer pore expansion is significantly reduced compared to the ATP-bound structure. Our data further reveal that the inner and outer ends of adjacent pore-lining helices come closer during opening, likely through a hinge-bending motion. These results provide new insight into the gating mechanism of P2X receptors and establish a versatile strategy applicable to other membrane proteins.