Characterization of an inducible aryl hydrocarbon receptor-like protein in rat liver.
The individual pretreatment of Sprague-Dawley rats with either 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) or 2,2',4,4',5,5'-hexachlorobiphenyl (HCB) has been previously shown to result in the "induction" of [3H]TCDD specific binding activity in hepatic tissue. In the present work, the coadministration of TCDD and HCB increased the concentration of hepatic proteins capable of binding [3H]TCDD specifically by at least 2-3-fold. This increase was shown not to be the result of activation, by HCB, of a form of the receptor having low affinity toward [3H]TCDD into a form with high affinity. Kinetic analysis of the time course of binding of [3H]TCDD to induced cytosol was consistent with the presence of an "inducible" binding protein in addition to the "constitutive" aryl hydrocarbon (Ah) receptor present in cytosol from untreated animals. The liganded ([3H]TCDD) form of the inducible binding component lost its ligand much faster than the liganded form of the constitutive Ah receptor at 37 degrees C; apparent first order rate constants for loss of [3H]TCDD were 0.55 min-1 and less than 0.0024 min-1, respectively. Conversely, the unliganded form of the induced binding component was slightly more stable (approximately 2-fold) toward thermal inactivation than the unbound constitutive Ah receptor. The [3H]TCDD-bound protein(s) in uninduced and induced cytosols behaved identically in a sucrose gradient; 8.7-8.9 S in the absence of salt, shifted to 5.5 S by 0.4 M KCl. They were also indistinguishable by gel permeation chromatography, and by photoaffinity labeling their TCDD-binding subunits, approximate molecular weights 105,000. These results show the hepatic TCDD-binding protein(s) induced upon pretreatment of Sprague-Dawley rats with TCDD/HCB to be kinetically distinct from the Ah receptor, but structurally very similar.
Landers
J P
JP
Department of Chemistry and Biochemistry, University of Guelph, Ontario, Canada.
Winhall
M J
MJ
McCready
T L
TL
Sanders
D A
DA
Rasper
D
D
Nakai
J S
JS
Bunce
N J
NJ
D000345:Affinity Labels; D000818:Animals; D001835:Body Weight; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006838:Hydrocarbons / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000737:chemistry*; D008297:Male; D009929:Organ Size; D000072317:Polychlorinated Dibenzodioxins / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D018336:Receptors, Aryl Hydrocarbon; D011955:Receptors, Drug / Q000737:chemistry*; D013696:Temperature; D013950:Thymus Gland / Q000187:drug effects
D000345:Affinity Labels; D006838:Hydrocarbons; D000072317:Polychlorinated Dibenzodioxins; D018336:Receptors, Aryl Hydrocarbon; D011955:Receptors, Drug