Androgen effects on the solubility and conformational change of the androgen receptor in baculovirus expression system.
To purify the androgen receptor (AR) efficiently from baculovirus expression system, we fused 6 histidine residues with the N-terminal domain of AR as a tag to specifically bind to Ni+2-affinity column. Our data indicated that adding androgen can increase the binding capacity of his-tag AR to the Ni+2-affinity column, and this increased binding capacity of AR could be due to the exposure of histidine residues of N-terminal domain induced by androgen. The androgen-enhanced binding to Ni+2-column also correlated with the increasing solubility of AR. Electrophoretic mobility shift assay further indicated that only purified AR could interact with androgen response element. Together, our data suggest that the binding of androgen to the hormone binding domain of AR may result in the conformational change of the N-terminal domain of AR and increase the hydrophilic property of AR.
Wang
C
C
Department of Pathology, University of Rochester, NY, USA.
Young
W J
WJ
Chang
C
C
D000728:Androgens / Q000494:pharmacology*; D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002460:Cell Line; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005786:Gene Expression Regulation / Q000187:drug effects*; D005822:Genetic Vectors; D006801:Humans; D011487:Protein Conformation / Q000187:drug effects; D011944:Receptors, Androgen / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012995:Solubility / Q000187:drug effects; D018411:Spodoptera
D000728:Androgens; D011944:Receptors, Androgen; D011994:Recombinant Proteins