Agonist binding evokes extensive conformational changes in the extracellular domain of the ATP-gated human P2X1 receptor ion channel

Roberts, J. A., Allsopp, R. C., El Ajouz, S., Vial, C., Schmid, R., Young, Mark Thomas ORCID: https://orcid.org/0000-0002-9615-9002 and Evans, R. J. 2012. Agonist binding evokes extensive conformational changes in the extracellular domain of the ATP-gated human P2X1 receptor ion channel. Proceedings of the National Academy of Sciences 109 (12) , pp. 4663-4667. 10.1073/pnas.1201872109

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Abstract

P2X receptors for ATP have a wide range of physiological roles and comprise a structurally distinct family of ligand-gated trimeric ion channels. The crystal structure of a P2X4 receptor, in combination with mutagenesis studies, has provided a model of the intersubunit ATP-binding sites and identified an extracellular lateral portal, adjacent to the membrane, that funnels ions to the channel pore. However, little is known about the extent of ATP-induced conformational changes in the extracellular domain of the receptor. To address this issue, we have used MTSEA-biotinylation (N-Biotinoylaminoethyl methanethiosulfonate) to show ATP-sensitive accessibility of cysteine mutants at the human P2X1 receptor. Mapping these data to a P2X1 receptor homology model identifies significant conformational rearrangement. Electron microscopy of purified P2X1 receptors showed marked changes in structure on ATP binding, and introducing disulphide bonds between adjacent subunits to restrict intersubunit movements inhibited channel function. These results are consistent with agonist-induced rotation of the propeller-head domain of the receptor, sliding of adjacent subunits leading to restricted access to the upper vestibule, movement in the ion conducting lateral portals, and gating of the channel pore.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Biosciences
Subjects:	Q Science > QH Natural history > QH301 Biology
Uncontrolled Keywords:	purinergic receptor; cysteine-scanning
Publisher:	National Academy of Sciences
ISSN:	0027-8424
Date of First Compliant Deposit:	30 March 2016
Last Modified:	17 May 2023 00:34
URI:	https://orca.cardiff.ac.uk/id/eprint/32221

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