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How Structural Biology Has Directly Impacted Our Understanding of P2X Receptor Function and Gating

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The P2X7 Receptor

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2510))

Abstract

P2X receptors are ATP-gated ion channels expressed in a wide variety of eukaryotic cells. They play key roles in diverse processes such as platelet activation, smooth muscle contraction, synaptic transmission, nociception, cell proliferation, and inflammation making this receptor family an important pharmacological target. Structures of P2X receptors solved by X-ray crystallography have been instrumental in helping to define mechanisms of molecular P2X receptor function. In 2009, the first X-ray structure of the P2X4 receptor subtype confirmed a trimeric stoichiometry and revealed the overall architecture of the functional ion channel. Subsequent X-ray structures have provided the molecular details to define the orthosteric ATP binding pocket, the orthosteric antagonist binding pocket, an allosteric antagonist binding pocket, and the pore architecture in each of the major conformational states of the receptor gating cycle. Moreover, the unique gating mechanism by which P2X receptor subtypes desensitize at differing rates, referred to as the helical recoil model of receptor desensitization, was discovered directly from X-ray structures of the P2X3 receptor. However, structures of P2X receptors solved by X-ray crystallography have only been able to provide limited information on the cytoplasmic domain of this receptor family, as this domain was always truncated to varying degrees in order to facilitate crystallization. Because the P2X7 receptor subtype has a significantly larger cytoplasmic domain that has been shown to be necessary for its ability to initiate apoptosis, an absence of structural information on the P2X7 receptor cytoplasmic domain has limited our understanding of its complex signaling pathways as well as its unusual ability to remain open without undergoing desensitization. This absence of cytoplasmic structural information for P2X7 receptors was recently overcome when the first full-length P2X7 receptor structures were solved by single-particle cryogenic electron microscopy. These structures finally provide insight into the large and unique P2X7 receptor cytoplasmic domain and revealed two novel structural elements and several surprising findings: first, a cytoplasmic structural element called the cytoplasmic ballast was identified that contains a dinuclear zinc ion complex and a high affinity guanosine nucleotide binding site and second, a palmitoylated membrane proximal structural element called the C-cys anchor was identified which prevents P2X7 receptor desensitization. This chapter will highlight the major structural and functional aspects of P2X receptors discovered through structural biology, with a key emphasis on the most recent cryogenic electron microscopy structures of the full-length, wild-type P2X7 receptor.

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Acknowledgments

I would like to thank Dr. Michael Godsey and Adam Oken for feedback on the manuscript. I would also like to thank Lori Vaskalis and Alanna McCarthy for help with the figures.

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Authors and Affiliations

  1. Department of Chemical Physiology and Biochemistry, Oregon Health & Science University, Portland, OR, USA

    Steven E. Mansoor

  2. Knight Cardiovascular Institute, Oregon Health & Science University, Portland, OR, USA

    Steven E. Mansoor

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Correspondence to Steven E. Mansoor .

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  1. Walther Straub Institute of Pharmacology and Toxicology, Ludwig-Maximilians-Universität München, Munich, Germany

    Annette Nicke

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Mansoor, S.E. (2022). How Structural Biology Has Directly Impacted Our Understanding of P2X Receptor Function and Gating. In: Nicke, A. (eds) The P2X7 Receptor. Methods in Molecular Biology, vol 2510. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2384-8_1

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  • DOI: https://doi.org/10.1007/978-1-0716-2384-8_1

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2383-1

  • Online ISBN: 978-1-0716-2384-8

  • eBook Packages: Springer Protocols

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