Characterization by photoaffinity labeling of a steroid binding protein in rat liver plasma membrane.
The mechanism of steroid uptake by the cell remains controversial. [3H]R5020 was utilized to characterize by photoaffinity labeling the steroid binding site in plasma membrane. This binding was saturable, reversible and had one type of binding site (Kd = 33 +/- 4 nM, Bmax = 32 +/- 2 pmol/mg). [3H]R5020 could be prevented from binding by a variety of steroids (cortisol, progesterone, deoxycorticosterone, and levonorgestrel); estradiol did not have affinity for this binding site. The kinetics of R5020 photoactivation was time dependent and saturable. SDS-PAGE showed a specific band which corresponded to a 53-kDa peptide. The sucrose density gradient analysis has revealed the existence of a protein with a sedimentation coefficient of 3.6 +/- 0.2 S. This polypeptide shows different characteristics than cytosolic steroid receptor or serum steroid binding proteins. This binding protein could correspond to the steroid binding site previously found in the plasma membrane.
Ibarrola
I
I
Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad del País Vasco/Euskal Herriko, Unibertsitatea, Bilbao, Spain.
Alejandro
A
A
Marino
A
A
Sancho
M J
MJ
Macarulla
J M
JM
Trueba
M
M
D000345:Affinity Labels; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D002462:Cell Membrane / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D003900:Desoxycorticosterone / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006854:Hydrocortisone / Q000378:metabolism; D016912:Levonorgestrel / Q000378:metabolism; D008099:Liver / Q000378:metabolism*; D008297:Male; D011374:Progesterone / Q000378:metabolism; D011397:Promegestone / Q000378:metabolism* / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D011980:Receptors, Progesterone / Q000378:metabolism*
D000345:Affinity Labels; D011980:Receptors, Progesterone; D003900:Desoxycorticosterone; D011374:Progesterone; D016912:Levonorgestrel; D011397:Promegestone; D006854:Hydrocortisone