6533b825fe1ef96bd1281dac

RESEARCH PRODUCT

Synthesis of biotin-labelled dexamethasone derivatives. Novel hormone-affinity probes.

Hans-jörg GrillIrmgard KöhlerKunhard PollowA. HeubnerBernhard Manz

subject

medicine.medical_treatmentBiotinBiochemistryBinding CompetitiveChromatography AffinityDexamethasoneSteroidchemistry.chemical_compoundGlucocorticoid receptorCytosolReceptors GlucocorticoidAffinity chromatographyBiotinCadaverinemedicineHumansBinding siteReceptorPolyacrylamide gel electrophoresisChromatographyBinding SitesChemistrySplenic NeoplasmsAffinity LabelsSteroid hormoneBiochemistryElectrophoresis Polyacrylamide Gel

description

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.

10.1111/j.1432-1033.1983.tb07266.xhttps://pubmed.ncbi.nlm.nih.gov/6832154