Open AccessSynthesis of Biotin‐Labelled Dexamethasone Derivatives
Author(s) -
MANZ Bernhard,
HEUBNER Arnulf,
KÖHLER Irmgard,
GRILL HansJörg,
POLLOW Kunhard
Publication year - 1983
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1983.tb07266.x
Subject(s) - sepharose , glucocorticoid receptor , avidin , chemistry , receptor , affinity chromatography , chromatography , nuclear receptor coactivator 2 , steroid , elution , biotin , glucocorticoid , biochemistry , nuclear receptor , hormone , biology , endocrinology , enzyme , transcription factor , gene
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‐16α‐methyl‐11β,17‐dihydroxy‐1,4‐androstadiene‐3‐one‐17β‐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 thousandfold can be obtained with good yield.