Open AccessSynthesis of a New Disulfide Affinity Adsorbent for Purification of Human Uterine Progesterone Receptor
Author(s) -
MANZ Bernhard,
GRILL HansJörg,
KÖHLER Irmgard,
HEUBNER Arnulf,
POLLOW Kunhard
Publication year - 1982
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1982.tb06959.x
Subject(s) - disulfide bond , chemistry , receptor , polyacrylamide gel electrophoresis , ligand (biochemistry) , gel electrophoresis , affinity chromatography , chromatography , progesterone receptor , adsorption , cleavage (geology) , biochemistry , biology , enzyme , organic chemistry , cancer , estrogen receptor , breast cancer , genetics , paleontology , fracture (geology)
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 ( K d 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.