Expression of the human androgen receptor in eukaryotic cells using a recombinant adenovirus vector yields high levels of the soluble, functional receptor protein.
The androgen receptor (AR) and closely related members of the steroid receptor family have proven difficult to obtain in native form in large quantities. In the case of the human AR (hAR), high-level expression in prokaryotic or non-mammalian cells leads to the synthesis of a high proportion of non-binding, insoluble, or degraded forms of the receptor protein. To circumvent these difficulties, we have constructed a recombinant adenovirus that directs the expression of hAR under the control of a potent, constitutive promoter. Infection of eukaryotic cells with this recombinant virus leads to the synthesis of large quantities of the intact AR. In contrast to expression methods designed to direct the full-length AR in bacteria, yeast, and insect cells, AR expressed in mammalian cells using this adenoviral vector accumulates at high levels, retains many properties of the native AR, and is not rapidly proteolyzed. This method will prove useful for large-scale preparations of hAR for use in functional and structural studies.
Zoppi
S
S
Department of Internal Medicine, Division of Endocrinology and Metabolism, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75235-8857, USA.
Allman
D R
DR
Gerard
R D
RD
McPhaul
M J
MJ
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002460:Cell Line; D013196:Dihydrotestosterone / Q000378:metabolism; D004958:Estradiol / Q000378:metabolism; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors; D006801:Humans; D006854:Hydrocortisone / Q000378:metabolism; D008024:Ligands; D008156:Luciferases / Q000235:genetics; D009277:Nandrolone / Q000031:analogs & derivatives / Q000378:metabolism; D011374:Progesterone / Q000378:metabolism; D011944:Receptors, Androgen / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012995:Solubility
D008024:Ligands; D011944:Receptors, Androgen; D011994:Recombinant Proteins; D013196:Dihydrotestosterone; D011374:Progesterone; D004958:Estradiol; D009277:Nandrolone; C100075:mibolerone; D008156:Luciferases; D006854:Hydrocortisone