Direct Chromatographic Determination of Dissociation Rate Constants of Ligand-Receptor Complexes: Assessment of the Interaction of Noncompetitive Inhibitors with an Immobilized Nicotinic Acetylcholine
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- 作者：Ruin Moaddel ; Krzysztof Jozwiak ; Rika Yamaguchi ; and Irving W. Wainer
- 刊名：Analytical Chemistry
- 出版年：2005
- 出版时间：August 15, 2005
- 年：2005
- 卷：77
- 期：16
- 页码：5421 - 5426
- 全文大小：141K
- 年卷期：v.77,no.16(August 15, 2005)
- ISSN：1520-6882
文摘
A liquid chromatographic stationary phase containingimmobilized membranes from a cell line expressing the
3
4 subtype of the neuronal nicotinic acetylcholinereceptor (nAChR) has been used to assess dissociationrate constants (kd) of 12 noncompetitive inhibitor-nAChRcomplexes. The pharmacological effects of the noncompetitive inhibitors, expressed as percent recovery ofactivity at 7 min and 4 h postexposure to the inhibitor,were also determined. The results demonstrate that thekd values correlated with the pharmacological effect andthat this approach can be used to identify molecularstructures associated with differences in kd values. Themethod can be adapted for use with membrane-boundreceptors, ion channels, and transporters and representsa direct and facile technique for the assessment ofdissociation rate constants (kd) of ligand-receptor complexes.
34 subtype of the neuronal nicotinic acetylcholinereceptor (nAChR) has been used to assess dissociationrate constants (kd) of 12 noncompetitive inhibitor-nAChRcomplexes. The pharmacological effects of the noncompetitive inhibitors, expressed as percent recovery ofactivity at 7 min and 4 h postexposure to the inhibitor,were also determined. The results demonstrate that thekd values correlated with the pharmacological effect andthat this approach can be used to identify molecularstructures associated with differences in kd values. Themethod can be adapted for use with membrane-boundreceptors, ion channels, and transporters and representsa direct and facile technique for the assessment ofdissociation rate constants (kd) of ligand-receptor complexes.