Interaction of progesterone receptor with immobilized adenosine triphosphate.
Affinity chromatography has been used to study the binding of ATP to cyto-plasmic progesterone receptors of hen oviduct. A resin which selectively binds the receptor protein was prepared by linking ATP covalently to Sepharose 4B through a 6-carbon bridge of adipic acid dihydrazide. Receptor bound to the affinity resin was recovered in a single peak upon gradient elution with KCl (0.2-1 M) or ATP (0-0.1 M). While affinity chromatography was normally accomplished using the [3H]progesterone receptor complex, the hormone was not necessary for ATP binding under the conditions employed. The chromatography of crude receptor preparations allowed up to 100-fold purification with greater than 80% recovery of the receptor. The semipurified receptor appeared intact when analysed by sucrose gradient centrifugation, polyacrylamide gel electrophoresis, and DEAE-cellulose chromatography. The latter procedure separated the receptor into two components, A and B, both of which were capable of binding ATP. Although a specific biochemical role of ATP in hormone receptor action has not been demonstrated, the present studies support this possibility and, in addition, offer a convenient and reliable step for the purification of progesterone receptors.
D000255:Adenosine Triphosphate; D000818:Animals; D002499:Centrifugation, Density Gradient; D002645:Chickens; D002846:Chromatography, Affinity; D005260:Female; D010057:Oviducts / Q000378:metabolism; D011374:Progesterone / Q000378:metabolism; D011980:Receptors, Progesterone / Q000302:isolation & purification / Q000378:metabolism
D011980:Receptors, Progesterone; D011374:Progesterone; D000255:Adenosine Triphosphate