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Potentiation of a neuronal nicotinic receptor via pseudo-agonist site

  • Simone Mazzaferro
  • Isabel Bermudez
  • Steven M. SineEmail author
Original Article
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Abstract

Neuronal nicotinic receptors containing α4 and β2 subunits assemble in two pentameric stoichiometries, (α4)3(β2)2 and (α4)2(β2)3, each with distinct pharmacological signatures; (α4)3(β2)2 receptors are strongly potentiated by the drug NS9283, whereas (α4)2(β2)3 receptors are unaffected. Despite this stoichiometry-selective pharmacology, the molecular identity of the target for NS9283 remains elusive. Here, studying (α4)3(β2)2 receptors, we show that mutations at either the principal face of the β2 subunit or the complementary face of the α4 subunit prevent NS9283 potentiation of ACh-elicited single-channel currents, suggesting the drug targets the β2–α4 pseudo-agonist sites, the α4–α4 agonist site, or both sites. To distinguish among these possibilities, we generated concatemeric receptors with mutations at specified subunit interfaces, and monitored the ability of NS9283 to potentiate ACh-elicited single-channel currents. We find that a mutation at the principal face of the β2 subunit at either β2–α4 pseudo-agonist site suppresses potentiation, whereas mutation at the complementary face of the α4 subunit at the α4–α4 agonist site allows a significant potentiation. Thus, monitoring potentiation of single concatemeric receptor channels reveals that the β2–α4 pseudo-agonist sites are required for stoichiometry-selective drug action. Together with the recently determined structure of the (α4)3(β2)2 receptor, the findings have implications for structure-guided drug design.

Keywords

Neurotransmitters Nicotine addiction Protein interfaces PAM Ligand-gated ion channels 

Abbreviations

nAChR

Nicotinic acetylcholine receptor

ACh

Acetylcholine

Notes

Acknowledgements

This research was supported by NIH Grant NS31744 to SMS.

Author contributions

SMS, SM, and IB conceived and coordinated the research; SM performed the research; SM and SMS analyzed the results; SMS and SM wrote the paper. All authors edited and approved the final version.

Compliance with ethical standards

Conflict of interest

The authors declare no competing interests.

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Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  • Simone Mazzaferro
    • 1
  • Isabel Bermudez
    • 4
  • Steven M. Sine
    • 1
    • 2
    • 3
    Email author
  1. 1.Receptor Biology Laboratory, Department of Physiology and Biomedical EngineeringMayo Clinic College of MedicineRochesterUSA
  2. 2.Department of NeurologyMayo Clinic College of MedicineRochesterUSA
  3. 3.Department of Molecular Pharmacology and Experimental TherapeuticsMayo Clinic College of MedicineRochesterUSA
  4. 4.School of Life SciencesOxford Brookes UniversityOxfordUK

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