Progesterone-binding sites of the chick oviduct receptor. Presence of a weaker ligand site which is destroyed by phosphatase treatment.
Titration of chick progesterone receptor over a wide range of [3H]progesterone concentration (0.15 to 90 nM) shows two distinct types of binding sites in cytosol and in partially purified receptor samples prepared from oviducts of estrogenized chicks. The difference in affinity between the two sites (Kd = 1 nM; Kd = 25 nM) is sufficient to allow analysis by Scatchard plot methods. Ligand competition studies show that both sites have the same relative specificity for progesterone compared to other steroids. Both sites seem to be on the same receptor molecule as shown by their copurification and chromatographic properties. No cooperativity between the two sites has been detected in analysis using either rate kinetics or equilibrium methods. Thus, the function of the low affinity sites is not apparent at this time; it does not appear to function as a "helper" site which influences binding to the high affinity site previously described. The binding constant of the low affinity site is sufficiently strong to allow potential occupancy of these sites in vivo, at least at certain stages of the female reproductive cycle. The hormone-binding activity of the low affinity site can be destroyed after in vitro treatment with alkaline phosphatase, but the high affinity site remains functional under these conditions. Inhibitors of the enzyme block the inactivation. Furthermore, preliminary data in vivo suggest that estrogen administration to the animal can influence the relative titer of the low affinity sites.
D000469:Alkaline Phosphatase / Q000494:pharmacology*; D000818:Animals; D002645:Chickens; D003600:Cytosol / Q000378:metabolism; D005260:Female; D007700:Kinetics; D008024:Ligands; D008970:Molecular Weight; D010057:Oviducts / Q000378:metabolism*; D011374:Progesterone / Q000378:metabolism; D011980:Receptors, Progesterone / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D013379:Substrate Specificity
D008024:Ligands; D011980:Receptors, Progesterone; D011374:Progesterone; D000469:Alkaline Phosphatase