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PreprintCited by 0Adenosine and Purinergic SignalingIon channel regulation and functionReceptor Mechanisms and Signaling

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Structural insights into the orthosteric inhibition of P2X receptors by non-ATP-analog antagonists

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Sheng, Danqi, Yue, Chenxi, Jin, Fei, Wang, Yao, Ichikawa, Muneyoshi, Yu, Ye, Guo, Chang-Run, Hattori, Motoyuki·2024·eLife
DOI 10.7554/elife.92829.2

Preprint — published as 10.7554/elife.92829.3 · Paper page

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Abstract

Abstract P2X receptors are extracellular ATP-gated ion channels that form homo-or heterotrimers and consist of seven subtypes. They are expressed in various tissues, including neuronal and nonneuronal cells, and play critical roles in physiological processes such as neurotransmission, inflammation, pain, and cancer. As a result, P2X receptors have attracted considerable interest as drug targets, and various competitive inhibitors have been developed. However, although several P2X receptor structures from different subtypes have been reported, the limited structural information of P2X receptors in complex with competitive antagonists hampers the understanding of orthosteric inhibition, hindering the further design and optimization of those antagonists for drug discovery. Here, we determined the cryo-EM structures of the mammalian P2X7 receptor in complex with two classical competitive antagonists of pyridoxal-5’-phosphate derivatives, PPNDS and PPADS, at 3.3 and 3.6 Å resolution, respectively, and performed structure-based mutational analysis by patch-clamp recording as well as MD simulations. Our structures revealed the orthosteric site for PPADS/PPNDS, and structural comparison with the previously reported apo-and ATP-bound structures showed how PPADS/PPNDS binding inhibits the conformational changes associated with channel activation. In addition, structure-based mutational analysis identified key residues involved in the PPNDS sensitivity of P2X1 and P2X3, which are known to have higher affinity for PPADS/PPNDS than other P2X subtypes. Overall, our work provides structural insights into the orthosteric inhibition and subtype specificity of P2X receptors by the classical P2X antagonists, pyridoxal-5’-phosphate derivatives, thereby facilitating the rational design of novel competitive antagonists for P2X receptors.

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Metadata

Authors
Sheng, D., Yue, C., Jin, F., Wang, Y., Ichikawa, M., Yu, Y., Guo, C.-R., & Hattori, M.
Affiliations
  • Fudan University (CN)
  • China Pharmaceutical University (CN)
  • School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University
Corresponding author
  • Guo, Chang-Run
  • Hattori, Motoyuki
ORCID
Venue
eLife
Publisher
eLife Sciences Publications, Ltd
Published
2024-03-20
ISSN
2050-084X (ISSN-L)
DOI
10.7554/elife.92829.2
OpenAlex
W4392993795
Cited by
0
References
1
Type
posted content
Language
en
Open access
Licence
Locations
eLife (acceptedVersion, cc-by)
Funders
  • National Natural Science Foundation of China (32071234, 32271244, 32250610205)
  • Shanghai Municipal Education Commission (ZDSYS14005)
  • Japan Science and Technology Agency (10.52926/JPMJPR20E1)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Physiology › Adenosine and Purinergic Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ion channel regulation and function
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
Keywords
P2X receptors, P2X7 receptor, competitive antagonists, PPADS, ATP-gated ion channels, receptor structure, channel activation, subtype specificity, P2X1, P2X3 receptor, structure-based drug design

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