Cell membrane chromatography competitive binding analysis for characterization of α1A adrenoreceptor binding interactions
Cell membrane chromatography competitive binding analysis for characterization of α1A adrenoreceptor binding interactions
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DOI：
10.1007/s00216-011-5026-z
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发表时间：
2011-07-01
影响因子：
4.3
通讯作者：
He, Langchong
中科院分区：
文献类型：
--
作者：
Du, Hui;Ren, Jing;He, Langchong
建立了一种新的高表达α（1A）肾上腺素受体（alpha（1A）AR）的细胞膜色谱（CMC）法，用于表征α（1A）AR结合相互作用。使用稳定表达高水平α（1A）AR的HEK 293 α（1A）细胞系制备CMC模型中的固定相。应用HEK 293 alpha（1A）/CMC-offline-HPLC系统特异性识别与alpha（1A）AR相互作用的配体，从模型中获得的解离平衡常数（K-D）为坦索罗辛为（1.87 +/- 0.13）× 10（-6）M，5-甲基乌拉地尔为（2.86 +/- 0.20）x 10（-6）M，多沙唑嗪为（3.01 +/- 0.19）× 10（-6）M，特拉唑嗪、阿夫唑嗪和酚妥拉明分别为（3.44 +/- 0.19）× 10（-6）M、（3.50 +/- 0.21）× 10（-6）M和（7.57 +/- 0.31）× 10（-6）M。坦索罗辛与特拉唑嗪之间的竞争结合研究表明，两种药物在α（1A）AAR的共同结合位点上相互作用。然而，坦索罗辛和羟甲唑啉之间的情况并非如此。结果表明，CMC结合改良竞争结合法是一种快速、有效的药物-受体相互作用研究方法。
A new high alpha(1A) adrenoreceptor (alpha(1A)AR) expression cell membrane chromatography (CMC) method was developed for characterization of alpha(1A)AR binding interactions. HEK293 alpha(1A) cell line, which expresses stably high levels of alpha(1A)AR, was used to prepare the stationary phase in the CMC model. The HEK293 alpha(1A)/CMC-offline-HPLC system was applied to specifically recognize the ligands which interact with the alpha(1A)AR, and the dissociation equilibrium constants (K-D) obtained from the model were (1.87 +/- 0.13) x 10(-6) M for tamsulosin, (2.86 +/- 0.20) x 10(-6) M for 5-methylurapidil, (3.01 +/- 0.19) x 10(-6) M for doxazosin, (3.44 +/- 0.19) x 10(-6) M for terazosin, (3.50 +/- 0.21) x 10(-6) M for alfuzosin, and (7.57 +/- 0.31) x 10(-6) M for phentolamine, respectively. The competitive binding study between tamsulosin and terazosin indicated that the two drugs interacted at the common binding site of alpha(1A)AAR. However, that was not the case between tamsulosin and oxymetazoline. The results had a positive correlation with those from radioligand binding assay and indicated that the CMC method combined modified competitive binding could be a quick and efficient way for characterizing the drug-receptor interactions.