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Citation: DU Hui, LI Jingjing, FAN Fei, YANG Huige, LIU Pengming, ZHENG Yan. Quantitative analysis of allosteric effects for 5-hydroxytryptamine receptor by cell membrane chromatography[J]. Chinese Journal of Chromatography, ;2017, 35(2): 191-195. doi: 10.3724/SP.J.1123.2016.07029 shu

Quantitative analysis of allosteric effects for 5-hydroxytryptamine receptor by cell membrane chromatography

Xi'an Institute for Food and Drug Control, Xi'an 710054, China

Received Date: 24 July 2016

Fund Project: Scientific Project of Shaanxi Administration of Traditional Chinese Medicine (No. 13-ZY005).

A rat striatum cell membrane chromatography (CMC) method was developed for characterizing the allosteric effects for 5-hydroxytryptamine receptor subtype 1D (5-HT_1D). The cell membrane stationary phase prepared by rat striatum was applied to specifically recognize the mixed standard solution of nitrendipine, imperatorin and ligustilide. Imperatorin was pumped through a CMC column continuously as a competitor to evaluate the effect of the competitor's concentrations on the retention of ligustilide. A decrease in the retention factor of ligustilide was observed in the range of 4.62×10^-7-4.62×10^-5mol/L. The addition of imperatorin weakened the affinity that ligustilide binding to 5-HT_1D by 2.06 times, which suggested that imperatorin occupied certain binding sites of 5-HT_1D on the column thus hindering the association of ligustilide. For further competitive studies, the mobile phase containing ligustilide (5.26×10^-7-5.26×10^-5mol/L) was pumped through the column, and imperatorin was injected onto the column. The equilibrium dissociation constant was (4.15±0.12) μmol/L for ligustilide, which corresponded well with the previous studies. The results indicated that the CMC method could be a new way for quantitative analysis of allosteric effects for 5-hydroxytryptamine receptor.
- cell membrane chromatography (CMC),
- 5-hydroxytryptamine (5-HT) receptor,
- allosteric effects,
- imperatorin,
- ligustilide
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