title: Synthesis of a new disulfide affinity adsorbent for purification of human uterine progesterone receptor.
abstract: For purification of the human uterine progesterone receptor, an affinity adsorbent was synthesized in which the specific ligand (16 alpha-ethyl-3-oxo-19nor-androst-4-ene 17 beta-carboxylic acid) was bound to derivatized celulose using a disulfide-group-containing spacer. The purified receptor protein, isolated by reductive cleavage of the disulfide bond, bound the synthetic gestagen R5020 with high affinity (Kd 12.2 nmol/l). The affinity gel was highly efficient. A 24000-fold purification of progesterone receptor with a recovery of 40% could be achieved in a single step within 6 h. By means of dodecyl sulphate/polyacrylamide gel electrophoresis two main polypeptides with molecular weights of about 43000 and 108000 could be demonstrated.
mesh_terms: D000327:Adsorption; D001667:Binding, Competitive; D002499:Centrifugation, Density Gradient; D002846:Chromatography, Affinity / Q000379:methods; D003600:Cytosol / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D011282:Pregnenediones / Q000138:chemical synthesis*; D045167:Progesterone Congeners / Q000138:chemical synthesis*; D011980:Receptors, Progesterone / Q000302:isolation & purification*; D012117:Resins, Synthetic / Q000138:chemical synthesis*; D014599:Uterus / Q000378:metabolism*
chemical_list: D011282:Pregnenediones; D045167:Progesterone Congeners; D011980:Receptors, Progesterone; D012117:Resins, Synthetic; C036597:ORG 2058 affinity resin