title: Purification of a human progesterone receptor.
abstract: The cytoplasmic progesterone receptor form human uterus has been purified to apparent homogeneity by a combination of ammonium sulfate fractionation and affinity chromatography. Affinity resins prepared by conventional means were compared to those prepared by a modified method. The latter give more reproducible results. A consistent finding was that low capacity resins gave the highest fold purification of the receptor. The pure receptor sedimented at 3.6 S on sucrose density gradient centrifugation, was eluted as a single band by 0.2 M KCl from DEAE-cellulose, and migrated as a single band of molecular weight 42 000 on NaDodSO4-polyacrylamide gel electrophoresis. Molecular weight determinations, obtained from Strokes' radii and sucrose gradient centrifugation, the receptors' behavior on ion exchange resins, and hormone binding specificity were all similar to those of the receptor found in crude cytosol. When the crude cytosol receptor was photoaffinity labeled by using 3H-labeled 17,21-dimethyl-19-norpregna-4,9-diene-3,20-dione followed by NaDodSO4-polyacrylamide gel electrophoresis, only protein of Mr 42 000 was labeled. This is consistent with our previous findings that alkylation of the pure receptor using 11-deoxycorticosterone bromo[3H]acetate showed labeling of a single protein of Mr 42 000. These properties confirm that the identity and integrity of the receptor have been maintained throughout its purification.
mesh_terms: D003600:Cytosol / Q000378:metabolism; D003900:Desoxycorticosterone / Q000378:metabolism; D005260:Female; D006801:Humans; D007700:Kinetics; D011374:Progesterone / Q000378:metabolism*; D011980:Receptors, Progesterone / Q000302:isolation & purification* / Q000528:radiation effects; D014466:Ultraviolet Rays; D014599:Uterus / Q000378:metabolism*
chemical_list: D011980:Receptors, Progesterone; D003900:Desoxycorticosterone; D011374:Progesterone