Synthesis of a New Disulfide Affinity Adsorbent for Purification of Human Uterine Progesterone Receptor

Publisher Website
Google Scholar
Abstract
For purification of the human uterine progesterone receptor, an affinity adsorbent was synthesized in which the specific ligand (16.alpha.-ethyl-3-oxo-19-nor-androst-4-ene 17.beta.-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 [promegestone] with high affinity (Kd 12.2 nmol/l). The affinity gel was highly efficient. A 24,000-fold purification of progesterone receptor with a recovery of 40% could be achieved in a single step within 6 h. By means of dodecyl sulfate/polyacrylamide gel electrophoresis 2 main polypeptides with MW of .apprx. 43,000 and 108,000 could be demonstrated.