Efficient eukaryotic expression system for authentic human sex hormone-binding globulin.
Sex hormone-binding globulin (SHBG) is the main carrier for androgens and oestrogens in humans. It mediates the transport of steroid hormones in the circulation and testicular fluid, and regulates their bioavailability to steroid-responsive tissues. In addition, the protein interacts with membrane receptors expressed in target tissues. Binding to the receptors is suspected to facilitate the uptake of steroid hormones and/or elicit cellular signal transduction. The identity of the SHBG receptor has not yet been resolved, in part due to a lack of sufficient quantities of authentic SHBG for receptor purification and molecular characterization. We have successfully addressed this problem by establishing an episomal expression system in human embryonic kidney cells that produces 5 mg of fully active human SHBG per litre. The recombinant protein resembles native SHBG in terms of structure, glycosylation pattern and steroid-binding activity. Moreover, the protein interacts with plasma membranes in steroid target tissues, an activity not observed with SHBG from other recombinant expression systems. Thus our studies have removed an important obstacle to the further elucidation of the role SHBG plays in steroid hormone action.
Hilpert
J
J
Max-Delbrueck-Center for Molecular Medicine, Robert-Roessle-Strasse 10, D-13125 Berlin, Germany.
Vorum
H
H
Burmeister
R
R
Spoelgen
R
R
Grishkovskaya
I
I
Misselwitz
R
R
Nykjaer
A
A
Willnow
T E
TE
D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002942:Circular Dichroism; D003001:Cloning, Molecular; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D004717:Endometrium / Q000378:metabolism; D004822:Epididymis / Q000378:metabolism; D004958:Estradiol / Q000378:metabolism*; D005260:Female; D006031:Glycosylation; D006801:Humans; D006854:Hydrocortisone / Q000378:metabolism*; D007668:Kidney; D007700:Kinetics; D008297:Male; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012738:Sex Hormone-Binding Globulin / Q000235:genetics* / Q000378:metabolism; D013739:Testosterone / Q000378:metabolism
D017931:DNA Primers; D011994:Recombinant Proteins; D012738:Sex Hormone-Binding Globulin; D013739:Testosterone; D004958:Estradiol; D006854:Hydrocortisone