TY - JOUR
T1 - Cell membrane chromatography competitive binding analysis for characterization of α1A adrenoreceptor binding interactions
AU - Du, Hui
AU - Ren, Jing
AU - Wang, Sicen
AU - He, Langchong
PY - 2011/7
Y1 - 2011/7
N2 - A new high α1A adrenoreceptor (α1AAR) expression cell membrane chromatography (CMC) method was developed for characterization of α1AAR binding interactions. HEK293 α1A cell line, which expresses stably high levels of α1AAR, was used to prepare the stationary phase in the CMC model. The HEK293 α1A/CMC-offline-HPLC system was applied to specifically recognize the ligands which interact with the α 1AAR, and the dissociation equilibrium constants (K D) obtained from the model were (1.87±0.13)×10-6 M for tamsulosin, (2.86±0.20)×10-6 M for 5-methylurapidil, (3.01±0.19)×10-6 M for doxazosin, (3.44±0.19) ×10-6 M for terazosin, (3.50±0.21)×10-6 M for alfuzosin, and (7.57±0.31)×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 α1AAR. 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. [Figure not available: see fulltext.]
AB - A new high α1A adrenoreceptor (α1AAR) expression cell membrane chromatography (CMC) method was developed for characterization of α1AAR binding interactions. HEK293 α1A cell line, which expresses stably high levels of α1AAR, was used to prepare the stationary phase in the CMC model. The HEK293 α1A/CMC-offline-HPLC system was applied to specifically recognize the ligands which interact with the α 1AAR, and the dissociation equilibrium constants (K D) obtained from the model were (1.87±0.13)×10-6 M for tamsulosin, (2.86±0.20)×10-6 M for 5-methylurapidil, (3.01±0.19)×10-6 M for doxazosin, (3.44±0.19) ×10-6 M for terazosin, (3.50±0.21)×10-6 M for alfuzosin, and (7.57±0.31)×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 α1AAR. 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. [Figure not available: see fulltext.]
KW - Binding site
KW - Cell membrane chromatography
KW - Dissociation equilibrium constant
KW - HEK293 α adrenoreceptor
KW - Modified competitive binding
UR - https://www.scopus.com/pages/publications/79960643520
U2 - 10.1007/s00216-011-5026-z
DO - 10.1007/s00216-011-5026-z
M3 - 文章
C2 - 21544540
AN - SCOPUS:79960643520
SN - 1618-2642
VL - 400
SP - 3625
EP - 3633
JO - Analytical and Bioanalytical Chemistry
JF - Analytical and Bioanalytical Chemistry
IS - 10
ER -