Synthesis of a new disulfide affinity adsorbent for purification of human uterine progesterone receptor.
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.
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*
D011282:Pregnenediones; D045167:Progesterone Congeners; D011980:Receptors, Progesterone; D012117:Resins, Synthetic; C036597:ORG 2058 affinity resin