Displacement and nonlinear chromatographic techniques in the investigation of interaction of noncompetitive inhibitors with an immobilized alpha3beta4 nicotinic acetylcholine receptor liquid chromatographic stationary phase

Anal Chem. 2002 Sep 15;74(18):4618-24. doi: 10.1021/ac0202029.


A liquid chromatographic column containing immobilized alpha3beta4 nicotinic acetylcholine receptors (alpha3beta4-nAChRs) has been used to determine the equilibrium association constants (Ka), desorption rate constants (kd), and adsorption rate constants (ka) for the noncompetitive inhibitors: mecamylamine, ketamine, bupropion, and dextromethorphan. Displacement chromatography, with mecamylamine as the displacer, was used to verify that the four compounds bound to the same site on the immobilized alpha3beta4-nAChRs. Nonlinear chromatographic techniques were then utilized to calculate the Ka, ka, and kd values associated with the formation of the noncompetitive inhibitor-alpha3beta4-nAChR complexes. The ka values determined in this study ranged from 19.7 to 10.5 microM(-1) sec(-1), with a relative order of mecamylamine > dextromethorphan > or = ketamine > bupropion. The kd values determined in this study indicated that dextromethorphan-induced inhibition should produce a longer recovery time than the other three NCIs. This was consistent with results from a previous in vitro study. The data from this study indicate that the immobilized alpha3beta4-nAChR column and nonlinear chromatography can be used in the study of NCIs at the alpha3beta4-nAChR.

MeSH terms

  • Chromatography, Liquid / methods*
  • Nicotinic Antagonists / pharmacology*
  • Receptors, Nicotinic / chemistry*
  • Receptors, Nicotinic / drug effects


  • Nicotinic Antagonists
  • Receptors, Nicotinic