Moaddel Ruin, Jozwiak Krzysztof, Yamaguchi Rika, Wainer Irving W
Gerontology Research Center, National institute on Aging, National Institutes of Health, 5600 Nathan Shock Drive, Baltimore, MD 21224, USA.
Anal Chem. 2005 Aug 15;77(16):5421-6. doi: 10.1021/ac0504464.
A liquid chromatographic stationary phase containing immobilized membranes from a cell line expressing the alpha3beta4 subtype of the neuronal nicotinic acetylcholine receptor (nAChR) has been used to assess dissociation rate constants (kd) of 12 noncompetitive inhibitor-nAChR complexes. The pharmacological effects of the noncompetitive inhibitors, expressed as percent recovery of activity at 7 min and 4 h postexposure to the inhibitor, were also determined. The results demonstrate that the kd values correlated with the pharmacological effect and that this approach can be used to identify molecular structures associated with differences in kd values. The method can be adapted for use with membrane-bound receptors, ion channels, and transporters and represents a direct and facile technique for the assessment of dissociation rate constants (kd) of ligand-receptor complexes.
一种液相色谱固定相，含有来自表达神经元烟碱型乙酰胆碱受体（nAChR）α3β4亚型的细胞系的固定化膜，已被用于评估12种非竞争性抑制剂 - nAChR复合物的解离速率常数（kd）。还测定了非竞争性抑制剂的药理作用，以暴露于抑制剂后7分钟和4小时时活性的恢复百分比表示。结果表明，kd值与药理作用相关，并且这种方法可用于鉴定与kd值差异相关的分子结构。该方法可适用于膜结合受体、离子通道和转运蛋白，是一种评估配体 - 受体复合物解离速率常数（kd）的直接且简便的技术。
