Functional analysis of the purified glucocorticoid receptor

Jan-Ake Gustafsson, Jan Carlstedt-Duke, Örjan Wrange, Sam Okret, Ann Charlotte Wikström

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

Glucocorticoid-receptor complex (GR) has been purified from rat liver by differential affinity for DNA before and after activation, followed by ion-exchange chromatography. The purified GR has mot. wt 94,000 dalton. The protein contains three functional domains: (A) a steroid-binding domain; (B) a DNA-binding domain; and (C) a domain necessary for normal biological function. A second protein, with mol. wt 72,000 dalton, copurifies with the GR. This protein does not bind steroid, does not interact with antibodies raised against the GR and does not show the same susceptibility to limited proteolytic cleavage as the 94,000 dalton protein. Analysis of the specific interaction of the purified GR with the mouse mammary tumour virus gene, assayed by glycerol-gradient centrifugation, shows that one molecule of 94,000 dalton protein binds to each of the specific binding sites in the long terminal repeat region Analysis of the fractions from the glycerol gradients show that the 72,000 dalton protein is associated to the binding species (94,000 dalton receptor protein) in about equimolar amounts. Analysis of the molybdate-stabilized non-activated receptor complex using monoclonal antibodies raised against the 94,000 dalton receptor protein indicates that the molybdate-stabilized complex is a hetero-oligomer. The hetero-oligomer consists of only one molecule of the 94,000 dalton receptor protein in association with other non-steroid-binding proteins. GR, glucocorticoid-receptor complex; MTV, mouse mammary tumour virus; TA, triamcinolone acetonide; mol. wt, molecular weight, Rs,Stokes radius.

Original languageEnglish (US)
Pages (from-to)63-68
Number of pages6
JournalJournal of Steroid Biochemistry
Volume24
Issue number1
DOIs
StatePublished - Jan 1 1986

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology

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