The plasma membrane of epididymal epithelial cells has a specific receptor which binds to androgen-binding protein and sex steroid-binding protein.
The binding of [3H] delta 6-testosterone photoaffinity-labelled rat androgen-binding protein (rABP) has been studied in an enriched fraction of plasma membranes of epithelial epididymal cells in immature (15 days) and adult rats (40 days). The binding was maximal in less than 30 min and more rapid at 4 degrees C than at 34 degrees C. It was calcium and pH dependent. Scatchard plots of the binding data gave curvilinear plots with two types of binding sites corresponding to a K(ass1) of 18.2 nM-1 and K(ass2) of 1.6 nM-1 (2.2 x 10(11) sites/mg protein and 5.4 x 10(11) sites/mg protein, respectively). In adult rats, only one type of binding site was found, with a K(ass) of 3.7 nM-1 (4.5 x 10(11) sites/mg protein). The number of receptors was 5-fold lower in the cauda than in the caput of the epididymis. The pretreatment of the isolated intact cells with streptozotocin induced a 45% reduction of the binding. Only unlabelled rABP and hSBP (human sex steroid-binding protein) but not other proteins (lactotransferrin, serotransferrin, asialofetuine, fetuine and bovine serum albumin) competed with the labelled ligand to bind plasma membranes. The membrane fraction was solubilized by triton X-100. Its incubation with labelled rABP and hSBP provoked the elution of the tracer as an aggregate into the void volume fraction of superose 6B mini-gel filtration columns. Structural homology between hSBP and rABP could be responsible for the common behaviour of the steroid-carrier molecules for the ABP receptor of rat epididymal epithelial cells.
Felden
F
F
Laboratoire de Biochimie Nutritionnelle et INSERM U 308, Faculté de Médecine, Vandoeuvre Les Nancy, France.
Leheup
B
B
Fremont
S
S
Bouguerne
R
R
Egloff
M
M
Nicolas
J P
JP
Grignon
G
G
Gueant
J L
JL
D000367:Age Factors; D000727:Androgen-Binding Protein / Q000378:metabolism*; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D004822:Epididymis / Q000378:metabolism* / Q000648:ultrastructure; D004848:Epithelium / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008297:Male; D051381:Rats; D011956:Receptors, Cell Surface / Q000378:metabolism*; D012738:Sex Hormone-Binding Globulin / Q000378:metabolism*; D013311:Streptozocin / Q000494:pharmacology; D013696:Temperature; D013997:Time Factors
D000727:Androgen-Binding Protein; D011956:Receptors, Cell Surface; D012738:Sex Hormone-Binding Globulin; D013311:Streptozocin