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 (16α-ethyl-3-oxo-19nor-androst-4-ene 17β-carboxylic acid) was bound to derivatized cellulose 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 6h. By means of dodecyl sulphate/polyacrylamide gel electrophoresis two main polypeptides with molecular weights of about 43000 and 108000 could be demonstrated.