Frontal analysis of cell-membrane chromatography for determination of drug-alpha(1D) adrenergic receptor affinity.
The aim of the present study was to determine drug-alpha(1D) adrenergic receptor (AR) affinity by frontal analysis of cell-membrane chromatography (CMC). The cell-membrane stationary phase (CMSP) was prepared by immobilizing rat aorta cell membranes on porous silica, and the resulting CMSP was used to determine drug binding affinity to alpha(1D)-AR by frontal analysis. The CMSP of rat aorta was stable and reproducible. Relative binding affinities (dissociation constant, K(d)) were determined by frontal chromatography for prazosin (166.13+/-18.36 nmol), BMY7378 (537.40+/-30.84 nmol), phentolamine (646.92+/-23.17 nmol), 5-methylurapidil (725.66+/-25.48 nmol), oxymetazoline (910.56+/-40.62 nmol) and methoxamine (1299.27+/-51.73 nmol). These results were consistent with the affinity rank order and showed a good correlation with the affinity of the same compounds for the cloned alpha(1D)-AR subtype obtained from radioligand-binding assay. The study demonstrates that frontal analysis of CMC may be used for direct determination of drug-receptor binding interactions, and that CMC is an alternative reliable method to quantitatively study ligand-receptor interactions.
Zeng
Aiguo
A
School of Medicine, Xi'an Jiaotong University, Xi'an 710061, PR China.
Yuan
Bingxiang
B
Wang
Changhe
C
Yang
Guangde
G
He
Langchong
L
D058646:Adrenergic alpha-1 Receptor Agonists; D058668:Adrenergic alpha-1 Receptor Antagonists; D000316:Adrenergic alpha-Agonists / Q000032:analysis / Q000378:metabolism* / Q000494:pharmacology; D000317:Adrenergic alpha-Antagonists / Q000032:analysis / Q000378:metabolism* / Q000494:pharmacology; D000818:Animals; D001013:Aorta, Thoracic / Q000737:chemistry; D001665:Binding Sites; D002462:Cell Membrane / Q000737:chemistry; D002846:Chromatography, Affinity / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley; D018340:Receptors, Adrenergic, alpha-1 / Q000378:metabolism*
D058646:Adrenergic alpha-1 Receptor Agonists; D058668:Adrenergic alpha-1 Receptor Antagonists; D000316:Adrenergic alpha-Agonists; D000317:Adrenergic alpha-Antagonists; D018340:Receptors, Adrenergic, alpha-1