Chromatography studies on bio-affinity of nine ligands of alpha1-adrenoceptor to alpha1D subtypes overexpressed in cell membrane.
To improve selectivity and specificity of cell membrane chromatography (CMC), the chromatography affinities of nine ligands of alpha1-adrenergic receptor(AR)to alpha1D-AR subtype were investigated. The human embryonic kidney (HEK) 293 cells expressed by cDNA of alpha1D-AR subtypes were cultured and cell membrane stationary phase (CMSP) was prepared. Then the interactions between ligands and alpha1D-AR in CMSP were investigated using CMC. The affinity rank order to alpha1D-AR subtype obtained from CMC for the nine alpha1-adrenoceceptor ligands is: prazosin, BMY7378, phentolamine, oxymetazoline, 5-methylurapidil, norepinephrine, phenylephrine, methoxamine, RS-17053. The affinity rank order is similar and correlates well with that obtained from others' radioligand binding assays (RBA). CMSP prepared by transfected HEK293 cells with alpha1-D-adrenoceptor cDNA and CMC method could be used to evaluate affinities of -drug-receptor and drug-receptor subtypes and to screen drugs selective to alpha1D-AR.
Zhang
Dian
D
School of Medicine, Xi'an Jiaotong University, Xi'an 710061, China.
Yuan
Bingxiang
B
Deng
Xiuling
X
Yang
Guangde
G
He
Langchong
L
Zhang
Youyi
Y
Han
Qide
Q
D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D002846:Chromatography, Affinity; D004353:Drug Evaluation, Preclinical; D015870:Gene Expression; D006801:Humans; D008024:Ligands; D018340:Receptors, Adrenergic, alpha-1 / Q000145:classification* / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
D008024:Ligands; D018340:Receptors, Adrenergic, alpha-1