Agonist-induced transitions of the acetylcholine receptor

Robert H Fairclough, Mark A. Agius, Eswari Gudipati, Laura Silvian, Brent Hamaoka, Christopher C. Beltzner, Mike Y. Lin, Anthony R. Chuang, David P Richman

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

Anti-acetylcholine receptor (AChR) monoclonal antibody 383C binds to the β-hairpin loop α(187-199) of only one of the two Torpedo AChR α subunits. The loop recognized is associated with the α subunit corresponding to the high-affinity d-tubocurarine (dTC) binding site. Desensitization of the receptor with carbamylcholine completely blocks the binding of 383C. Mild reduction of AChR α subunit cys 192-193 disulfide with DTT and subsequent reaction with 5-iodoacetamidofluorescein label only the high-affinity dTC α subunit. Rhodamine-labeled α-bungarotoxin (R-Btx) binds to the unlabeled AChR α subunit as monitored by fluorescence resonance energy transfer between the fluorescein and rhodamine dyes. A 10-Å contraction of the distance between the dyes is observed following the addition of carbamylcholine. In a small angle X-ray diffraction experiment exploiting anomalous X-ray scattering from Tb(III) ions titrated into AChR Ca(II) binding sites, we find evidence for a change in the Tb(III) ion distribution in the region of the ion channel following addition of carbamylcholine to the AChR. The carbamylcholine-induced loss of the 383C epitope, the 10-Å contraction of the β-hairpin loop, and the loss of multivalent cations from the channel likely represent the first molecular transitions leading to AChR channel opening.

Original languageEnglish (US)
Pages (from-to)101-113
Number of pages13
JournalAnnals of the New York Academy of Sciences
Volume998
DOIs
StatePublished - 2003

Keywords

  • Acetylcholine receptor (AChR)
  • Agonist
  • Carbamylcholine
  • D-tubocurarine (dTC)
  • ELISA
  • FRET
  • X-ray diffraction
  • X-ray scattering

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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  • Cite this

    Fairclough, R. H., Agius, M. A., Gudipati, E., Silvian, L., Hamaoka, B., Beltzner, C. C., Lin, M. Y., Chuang, A. R., & Richman, D. P. (2003). Agonist-induced transitions of the acetylcholine receptor. Annals of the New York Academy of Sciences, 998, 101-113. https://doi.org/10.1196/annals.1254.012