Engineered site-directed labeling of nicotinic acetylcholine receptors using reactive epibatidine derivatives: appraisal of epibatidine-docking models in neuronal and muscular receptors.

Conference Paper

We developed an engineered site-directed labeling method (Foucaud et al., 2001) to investigate ligand receptor interactions on the acetylcholine (ACh)- binding site of nicotinic acetylcholine receptors (nAChRs). The method uses cysteine receptor mutants, together with cysteine-reactive ligand analogs, to generate a site-directed covalent reaction within the binding site. We selected epibatidine (EPB) as a prototypical ligand, acting at all types of nAChRs with sufficient affinity to allow this study. Accordingly, we synthesized three cysteine-reactive derivatives, all modified at the C-3 of the pyridine ring of the alkaloid with NCS; -NHCOCH2Cl, and -CH2Cl groups, respectively (Fig. 1). The binding properties have been established on rat brain, alpha7-5HT3 chimera, and Torpedo membranes, respectively, whereas the functional properties were tested on alpha4beta2 and alpha7 receptor expressed in oocytes and Cys-less muscular receptor expressed in HEK cells (Sakr et al., 2005).

Full Text

Duke Authors

Cited Authors

  • Sakr, E; Kotzyba-Hibert, F; Grande, J; Hovius, R; Vogel, H; Bertrand, S; Bertrand, D; Goeldner, M

Published Date

  • 2006

Published In

Volume / Issue

  • 30 / 1-2

Start / End Page

  • 35 - 36

PubMed ID

  • 17192618

International Standard Serial Number (ISSN)

  • 0895-8696

Digital Object Identifier (DOI)

  • 10.1385/JMN:30:1:35

Conference Location

  • United States