Using the protein chip to screen agonists and antagonists of the androgen receptor.
Based on its important physical and pathological function, the androgen receptor (AR) is regarded as a significant drug target. In this report, the authors describe a novel strategy of protein chip technology to screen agonists and antagonists of AR. First, the AR ligand binding domain (AR-LBD) was expressed in Escherichia coli, purified, and then immobilized on a silane-polysaccharide surface of a protein chip. Second, the affinities of methyltestosterone (MT) and fluorescent-labeled testosterone for the AR-LBD protein chip were determined. Third, a converse strategy of the protein chip was tested to evaluate its reliability as a drug screening method. Fourth, a 10,067-compound library was screened to find new ligands of AR. From the results, the K(d) of testosterone and the IC(50) of MT are consistent with the literature (0.61 vs. 0.49 nM 2.88 vs. 3.90 nM, respectively). The Z' factor of the high-throughput screening (HTS) method was 0.76, which meets the requirement of drug screening (>0.4). Finally, 3 active ligands of AR were identified with their IC( 50) values of 3.63, 2.19, and 1.71 microM, respectively. In summary, the novel strategy of the AR-LBD protein chip was suitable for HTS at the molecular level.
Zhou
Yong
Y
National Center of Pharmaceutical Screening, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China, Ottawa Health Research Institute, Faculty of Medicine, University of Ottawa, Ottawa, Canada.
Liu
Ailin
A
Wang
Wei
W
Du
Guanhua
G
D059002:Androgen Receptor Antagonists; D000728:Androgens; D000818:Animals; D004353:Drug Evaluation, Preclinical / Q000295:instrumentation / Q000379:methods*; D016650:Fluorescein-5-isothiocyanate; D008297:Male; D008777:Methyltestosterone / Q000378:metabolism; D040081:Protein Array Analysis; D017434:Protein Structure, Tertiary; D051381:Rats; D017208:Rats, Wistar; D011944:Receptors, Androgen / Q000737:chemistry / Q000302:isolation & purification
D059002:Androgen Receptor Antagonists; D000728:Androgens; D011944:Receptors, Androgen; D016650:Fluorescein-5-isothiocyanate; D008777:Methyltestosterone