TY - JOUR
T1 - High-affinity binding of 2,3,7,8-tetrachlorodibenzo-p-dioxin in cell nuclei from rat liver
AU - Poellinger, Lorenz
AU - Kurl, Rabinder Nath
AU - Lund, Johan
AU - Gillner, Mikael
AU - Carlstedt-Duke, Jan
AU - Högberg, Bertil
AU - Gustafsson, Jan Ake
N1 - Funding Information:
This work was supported by grants from the Swedish Cancer Society (Riksf6reningen mot Cancer), Leo Research Foundation and the Swedish Tobacco Company (Grant No. 7910). R.N.K. and M.G. are grateful to Ekhagastiftelsen for Fellowships. R.N.K. is also a recipient of a Council of Europe Fellowship from the Swedish Institute. J.C.-D. is a recipient of a Research Fellowship from the Swedish Cancer Society.
Copyright:
Copyright 2014 Elsevier B.V., All rights reserved.
PY - 1982/2/25
Y1 - 1982/2/25
N2 - 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.
AB - 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.
KW - (Rat liver)
KW - Nuclear binding
KW - Tetrachlorodibenzo-p-dioxin
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U2 - 10.1016/0304-4165(82)90162-3
DO - 10.1016/0304-4165(82)90162-3
M3 - Article
C2 - 6277389
AN - SCOPUS:0020030658
SN - 0304-4165
VL - 714
SP - 516
EP - 523
JO - BBA - General Subjects
JF - BBA - General Subjects
IS - 3
ER -