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Molecular Pharmacology

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Research ArticleArticle

Alternate Stoichiometries of α4β2 Nicotinic Acetylcholine Receptors

Mark E. Nelson, Alexander Kuryatov, Catherine H. Choi, Yan Zhou and Jon Lindstrom
Molecular Pharmacology February 2003, 63 (2) 332-341; DOI: https://doi.org/10.1124/mol.63.2.332
Mark E. Nelson
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Alexander Kuryatov
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Catherine H. Choi
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Yan Zhou
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Jon Lindstrom
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Abstract

Two functional types of nicotinic acetylcholine receptors (nAChRs) are expressed when human embryonic kidney cells are permanently transfected with equal amounts of human α4 and β2 subunit cDNAs. Most (82%) of these nAChRs exhibit an EC50of 74 ± 6 μM for ACh, a much lower sensitivity than the remaining fraction (EC50 of 0.7 ± 0.4 μM) or than expected from expression of equal amounts of α4 and β2 mRNAs inXenopus laevis oocytes. We have found three conditions that can increase the number of nAChRs with high sensitivity to activation. These are: 1) transient transfection with additional β2 subunits, 2) overnight incubation in nicotine, or 3) overnight culture at 29°C. Using metabolic labeling with [35S]methionine to measure subunit stoichiometry, we found that the majority of nAChRs had a stoichiometry of (α4)3(β2)2. Overnight treatment with nicotine increased the number of nAChRs and increased the proportion of the (α4)2(β2)3stoichiometry. Alternate α4β2 nAChR stoichiometries with distinct functional properties raise the possibility for an interesting mode of synaptic regulation for nicotinic signaling in the mammalian brain.

Footnotes

  • This work supported by grant NS11323 from the National Institutes of Health and the Smokeless Tobacco Research Council (to J.L.).

  • Abbreviations:
    nAChR
    nicotinic acetylcholine receptor
    HEK
    human embryonic kidney
    mAb
    monoclonal antibody
    PBS
    phosphate-buffered saline
    DMEM
    Dulbecco's modified Eagle's medium
    PAGE
    polyacrylamide gel electrophoresis
    DMPP
    1,1-dimethyl-4-phenylpiperazinium
    • Received July 30, 2002.
    • Accepted October 30, 2002.
  • The American Society for Pharmacology and Experimental Therapeutics
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Molecular Pharmacology: 63 (2)
Molecular Pharmacology
Vol. 63, Issue 2
1 Feb 2003
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Research ArticleArticle

Alternate Stoichiometries of α4β2 Nicotinic Acetylcholine Receptors

Mark E. Nelson, Alexander Kuryatov, Catherine H. Choi, Yan Zhou and Jon Lindstrom
Molecular Pharmacology February 1, 2003, 63 (2) 332-341; DOI: https://doi.org/10.1124/mol.63.2.332

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Research ArticleArticle

Alternate Stoichiometries of α4β2 Nicotinic Acetylcholine Receptors

Mark E. Nelson, Alexander Kuryatov, Catherine H. Choi, Yan Zhou and Jon Lindstrom
Molecular Pharmacology February 1, 2003, 63 (2) 332-341; DOI: https://doi.org/10.1124/mol.63.2.332
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