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An Angstrom Scale Interaction between Plasma Membrane ATP-Gated P2X₂ and α₄β₂ Nicotinic Channels Measured with Fluorescence Resonance Energy Transfer and Total Internal Reflection Fluorescence Microscopy

Khakh, Baljit S. and Fisher, James A. and Nashmi, Raad and Bowser, David N. and Lester, Henry A. (2005) An Angstrom Scale Interaction between Plasma Membrane ATP-Gated P2X₂ and α₄β₂ Nicotinic Channels Measured with Fluorescence Resonance Energy Transfer and Total Internal Reflection Fluorescence Microscopy. Journal of Neuroscience, 25 (29). pp. 6911-6920. ISSN 0270-6474. PMCID PMC6725363. doi:10.1523/jneurosci.0561-05.2005. https://resolver.caltech.edu/CaltechAUTHORS:20200417-142718484

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Abstract

Structurally distinct nicotinic and P2X channels interact functionally, such that coactivation results in cross-inhibition of one or both channel types. It is hypothesized, but not yet proven, that nicotinic and P2X channels interact at the plasma membrane. Here, we show that plasma membrane α₄β₂ nicotinic and P2X₂ channels form a molecular scale partnership and also influence each other when coactivated, resulting in nonadditive cross-inhibitory responses. Total internal reflection fluorescence and fluorescence resonance energy transfer microscopy between fluorescently labeled P2X₂ and α₄β₂ nicotinic channels demonstrated close spatial arrangement of the channels in human embryonic kidney cells and in hippocampal neuron membranes. The data suggest that P2X₂ and α₄β₂ channels may form a dimer, with the channels ∼80 Å apart. The measurements also show that P2X₂ subunits interact specifically and robustly with the β₂ subunits in α₄β₂ channels. The data provide direct evidence for the close spatial apposition of full-length P2X₂ and α₄β₂ channels within 100 nm of the plasma membrane of living cells.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1523/jneurosci.0561-05.2005DOIArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/pmc6725363PubMed CentralArticle
ORCID:
AuthorORCID
Lester, Henry A.0000-0002-5470-5255
Additional Information:© 2005 Society for Neuroscience. Received Feb. 10, 2005; revised June 13, 2005; accepted June 14, 2005. Research in our laboratories is supported by the Medical Research Council (MRC), the European Molecular Biology Organization, the Human Frontiers Science Program, and National Institutes of Health Grants NS11756 and DA17279. J.A.F. was supported by an MRC Studentship, R.N. was supported by a postdoctoral fellowship from the California Tobacco-Related Disease Research Program and the Elizabeth Ross Fund, and D.N.B. was supported in part by a European Molecular Biology Organization Fellowship. We thank Julian Revie for constructing some of the nicotinic ACh receptor cDNAs.
Funders:
Funding AgencyGrant Number
Medical Research Council (UK)UNSPECIFIED
European Molecular Biology Organization (EMBO)UNSPECIFIED
Human Frontier Science ProgramUNSPECIFIED
NIHNS11756
NIHDA17279
California Tobacco-Related Disease Research ProgramUNSPECIFIED
Elizabeth Ross FellowshipUNSPECIFIED
Subject Keywords:channel; cholinergic; purinergic; acetylcholine receptor; ACh; fluorescence microscopy; P2X
Issue or Number:29
PubMed Central ID:PMC6725363
DOI:10.1523/jneurosci.0561-05.2005
Record Number:CaltechAUTHORS:20200417-142718484
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20200417-142718484
Official Citation:An Angstrom Scale Interaction between Plasma Membrane ATP-Gated P2X2 and α4β2 Nicotinic Channels Measured with Fluorescence Resonance Energy Transfer and Total Internal Reflection Fluorescence Microscopy Baljit S. Khakh, James A. Fisher, Raad Nashmi, David N. Bowser, Henry A. Lester Journal of Neuroscience 20 July 2005, 25 (29) 6911-6920; DOI: 10.1523/JNEUROSCI.0561-05.2005
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:102638
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:17 Apr 2020 21:47
Last Modified:16 Nov 2021 18:14

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