Molecular shape, architecture, and size of P2X4 receptors determined using fluorescence resonance energy transfer and electron microscopy

Young, Mark Thomas ORCID: https://orcid.org/0000-0002-9615-9002, Fisher, J., Fountain, S. J., Ford, R. C., North, R. and Khakh, B. S. 2008. Molecular shape, architecture, and size of P2X4 receptors determined using fluorescence resonance energy transfer and electron microscopy. Journal of Biological Chemistry 283 (38) , pp. 26241-26251. 10.1074/jbc.M804458200

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Abstract

P2X receptors are ATP-gated nonselective cation channels with important physiological roles. However, their structures are poorly understood. Here, we analyzed the architecture of P2X receptors using fluorescence resonance energy transfer (FRET) microscopy and direct structure determination using electron microscopy. FRET efficiency measurements indicated that the distance between the C-terminal tails of P2X4 receptors was 5.6 nm. Single particle analysis of purified P2X4 receptors was used to determine the three-dimensional structure at a resolution of 21Å; the orientation of the particle with respect to the membrane was assigned by labeling the intracellular C termini with 1.8-nm gold particles and the carbohydrate-rich ectodomain with lectin. We found that human P2X4 is a globular torpedo-like molecule with an approximate volume of 270 nm3 and a compact propeller-shaped ectodomain. In this structure, the distance between the centers of the gold particles was 6.1 nm, which closely matches FRET data. Thus, our data provide the first views of the architecture, shape, and size of single P2X receptors, furthering our understanding of this important family of ligand-gated ion channels.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Biosciences
Subjects:	Q Science > Q Science (General)
Publisher:	American Society for Biochemistry and Molecular Biology
ISSN:	0021-9258
Date of First Compliant Deposit:	30 March 2016
Last Modified:	06 May 2023 05:21
URI:	https://orca.cardiff.ac.uk/id/eprint/22584

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