The size and/or configuration of the cycloalkane D' ring in pentacyclic progesterone derivatives are crucial for their high-affinity binding to a protein in addition to progesterone receptor in rat uterine cytosol.
[(3)H]labeled progesterone and a number of its 16alpha, 17alpha-cycloalkano derivatives with an additional three to six-membered D' ring were investigated for mutual competition and equilibrium binding to proteins from rat uterine cytosol. The interaction of all studied [(3)H]ligands with proteins was characterized by comparable affinity (K(d) in nM region) and apparent homogeneity in terms of affinity. At the same time, the concentrations of binding sites for ligands bearing 16alpha,17alpha cyclopentano, cyclohexano, or cyclohexeno substituents were several-fold higher than those for progesterone or 16alpha, 17alpha-cyclopropanoprogesterone. In mutual competition experiments, when [(3)H]progesterone or [(3)H]16alpha, 17alpha-cyclopropanoprogesterone were used, the curves of 'bound radioactivity-log of competitor concentration' for all compounds studied were parallel and corresponded to a model of 'one protein-two ligands.' However, when [(3)H]ligands with bulky 16alpha, 17alpha-substituents (with the possible exception of cyclohexene derivative) were used, competitive curves for various ligands had different appearances and fell into two groups. Parallel curves for derivatives with 5 or 6 carbons in D' ring described by a model of 'one protein-two ligands' formed the 1st group. The 2nd group comprised curves for progesterone or 16alpha, 17alpha-cyclopropanoprogesterone that had lower slopes and could be described by a model of 'two proteins-two ligands.' Taken together, the results suggest the presence in rat uterine cytosol, of a protein in addition to progesterone receptor capable of discriminating between ligands with no or small 16alpha, 17alpha-cycloalkano substituents and ligands with more bulky substituents.
Smirnov
A N
AN
Laboratory of Endocrinology, School of Biology, Lomonosov Moscow State University, Lenin Hills 1/12, Moscow, Russia. smirnov_an@hotmail.com
Pokrovskaya
E V
EV
Kogteva
G S
GS
Shevchenko
V P
VP
Levina
I S
IS
Kulikova
L E
LE
Kamernitzky
A V
AV
D000473:Alkanes / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D002850:Chromatography, Gel; D003600:Cytosol / Q000378:metabolism; D003907:Dexamethasone / Q000378:metabolism; D013196:Dihydrotestosterone / Q000378:metabolism; D004958:Estradiol / Q000378:metabolism; D005260:Female; D008297:Male; D011374:Progesterone / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D011944:Receptors, Androgen / Q000378:metabolism; D011960:Receptors, Estrogen / Q000378:metabolism; D011965:Receptors, Glucocorticoid / Q000378:metabolism; D011980:Receptors, Progesterone / Q000378:metabolism*; D013329:Structure-Activity Relationship; D014599:Uterus / Q000166:cytology / Q000378:metabolism
C113166:16,17-cyclopropanoprogesterone; D000473:Alkanes; D011506:Proteins; D011944:Receptors, Androgen; D011960:Receptors, Estrogen; D011965:Receptors, Glucocorticoid; D011980:Receptors, Progesterone; D013196:Dihydrotestosterone; C085261:pentarane A; D011374:Progesterone; D004958:Estradiol; C085262:pentarane B; D003907:Dexamethasone