Synthesis of biotin-labelled dexamethasone derivatives. Novel hormone-affinity probes.
A new, general methodology for 'sandwich' affinity chromatography of steroid hormone receptors is proposed, the part purification of the human spleen tumor glucocorticoid receptor is quoted as an illustration. 9-Fluoro-16 alpha-methyl-11 beta, 17-dihydroxy-1,4-androstadiene-3-one-17 beta-carboxylic acid was coupled to biotin using pentamethylenediamine (BioDex 1) as a spacer. The bifunctional derivative binds to glucocorticoid receptors and avidin-Sepharose and efficiently protects the glucocorticoid receptor against inactivation when previously added during homogenisation. We have standardized the capacity and optimum conditions for elution of receptor-BioDex-1 complexes which are bound to avidin-Sepharose. Receptor purification of several thousand fold can be obtained with good yield.
D000345:Affinity Labels / Q000138:chemical synthesis*; D001665:Binding Sites; D001667:Binding, Competitive; D001710:Biotin / Q000031:analogs & derivatives* / Q000138:chemical synthesis; D002103:Cadaverine / Q000138:chemical synthesis; D002846:Chromatography, Affinity; D003600:Cytosol / Q000378:metabolism; D003907:Dexamethasone / Q000031:analogs & derivatives* / Q000138:chemical synthesis; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D011965:Receptors, Glucocorticoid / Q000302:isolation & purification; D013160:Splenic Neoplasms / Q000378:metabolism
D000345:Affinity Labels; D011965:Receptors, Glucocorticoid; C028697:dexamethasone receptor; D001710:Biotin; D003907:Dexamethasone; D002103:Cadaverine