High-affinity binding of 2,3,7,8-tetrachlorodibenzo-p-dioxin in cell nuclei from rat liver

Lorenz Poellinger, Rabinder Nath Kurl, Johan Lund, Mikael Gillner, Jan Carlstedt-Duke, Bertil Högberg, Jan Ake Gustafsson

Research output: Contribution to journalArticle

29 Scopus citations

Abstract

The intranuclear binding of radioactive 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in rat liver has been studied both in vivo and in vitro. Following the intravenous administration of [1,6-3H]TCDD, a maximum uptake by cell nuclei could be observed at 2 h after injection with a concurrent decrease in the cytosolic uptake. Using linear sucrose density gradient centrifugation, dextran-coated charcoal adsorption assay, DEAE-Sepharose ion-exchange chromatography, competition, enzymatic and saturation studies, a high-affinity binding protein for TCDD in liver cell nuclei could be demonstrated both in vivo and after an exchange in vitro of intravenously administered unlabelled 2,3,7,8- tetrachlorodibenzofuran (TCDBF) for [3H]TCDD. Sucrose density gradient analysis showed a size of 4-5 S for both the cytosolic and nuclear TCDD binding entity. The specific binding of [3H]TCDD to nuclear components was heat labile and saturable and had an equilibrium dissociation constant of 1.05 nM. Based on a differential susceptibility to specific hydrolases, i.e. DNAase, RNAase, trypsin and pronase, the binding entity appears to be a 4-5 S salt-extractable protein.

Original languageEnglish (US)
Pages (from-to)516-523
Number of pages8
JournalBBA - General Subjects
Volume714
Issue number3
DOIs
StatePublished - Feb 25 1982

Keywords

  • (Rat liver)
  • Nuclear binding
  • Tetrachlorodibenzo-p-dioxin

ASJC Scopus subject areas

  • Molecular Biology
  • Biophysics
  • Biochemistry
  • Medicine(all)

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