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Alternative ligands for measurement and purification of ecdysteroid receptors in Drosophila K c cells
Author(s) -
Sage Becky A.,
Horn Denis H. S.,
Landon Thomas M.,
O'Connor John D.
Publication year - 1986
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/arch.940030705
Subject(s) - ecdysteroid , moiety , biology , cytosol , receptor , biochemistry , binding protein , stereochemistry , chemistry , enzyme , hormone , gene
An ecdysteroid‐binding protein has been demonstrated in nuclear and cytosolic extracts of cultured Drosophila cells (K c cells). Attempts to purify the binding moiety have been hampered because of the small number of binding sites (ca 1,000/cell) and sensitivity of the ecdysteroid binding moiety toward salt (dissociation at 150 mM). Recently 3α[ 3 H]kaladasterone and 3α[ 3 H]muristerone A have been synthesized. The binding kinetics of these two ecdysteroids are similar to ponasterone A. Photoaffinity labeling of the ecdysteroid receptor in Kc cells was attempted using the 3α[ 3 H]kaladasterone, and standard protein chromatographic techniques have been used to examine the 3α[ 3 H]muristerone A‐receptor complex, which is less sensitive to salt dissociation. Attempts to characterize the protein to which these ligands have been attached will be discussed.