Counterregulation of nuclear 3,5,3'-triiodo-L-thyronine (T3) binding by oxidized and reduced-nicotinamide adenine dinucleotide phosphates in the presence of cytosolic T3-binding protein in vitro.
The role of cytosolic T3-binding protein (CTBP) in the regulation of nuclear T3 binding was studied in vitro. Nuclear [125I]T3 binding was observed in the presence of 1.0 mM dithiothreitol (DTT). When the nuclei prepared from rat kidney were incubated with inactive form of CTBP which was also prepared from rat kidney, [125I]T3 binding to nuclei was not affected. When the nuclei were incubated with inactive form of CTBP in the presence of NADP, [125I]T3 binding to nuclei was increased, whereas binding was diminished when nuclei were incubated with CTBP in the presence of NADPH. The inactive form of CTBP was activated by NADPH. NADP also activated CTBP in the presence of DTT. Both active forms of CTBP were again inactivated by extraction with charcoal, and these inactive forms were reactivated by NADPH or by NADP and DTT, but not by NADP alone. Although the nuclei treated with 0.3 M NaCl lost the binding activity for [125I]T3 in the absence of NADP, the nuclei retained the binding activity for [125I]T3 in the presence of NADP and the inactive form of CTBP. Treatment of the nuclei with 0.5 M NaCl lost the binding activity for [125I]T3 not only in the absence but also in the presence of NADP and CTBP. These results suggested that NADP and NADPH play roles as counterregulatory factors for nuclear T3 binding in the presence of CTBP. Further, it was speculated that binding sites for the T3-CTBP complex, which is generated in the presence of NADP and DTT, are present in nuclei, and that binding sites for the complex are different from nuclear T3 receptors.
Hashizume
K
K
Department of Gerontology, Endocrinology, and Metabolism, Shinshu University School of Medicine, Matsumoto, Japan.
Miyamoto
T
T
Yamauchi
K
K
Ichikawa
K
K
Kobayashi
M
M
Ohtsuka
H
H
Sakurai
A
A
Suzuki
S
S
Yamada
T
T
D000818:Animals; D002352:Carrier Proteins / Q000032:analysis / Q000378:metabolism* / Q000494:pharmacology / Q000502:physiology; D002467:Cell Nucleus / Q000032:analysis / Q000378:metabolism; D003600:Cytosol / Q000032:analysis / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology; D007457:Iodine Radioisotopes; D008297:Male; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism* / Q000494:pharmacology; D009249:NADP / Q000378:metabolism*; D010084:Oxidation-Reduction; D051381:Rats; D011919:Rats, Inbred Strains; D012965:Sodium Chloride / Q000494:pharmacology; D013963:Thyroid Hormones; D013997:Time Factors; D014284:Triiodothyronine / Q000032:analysis / Q000378:metabolism*; D000097810:Thyroid Hormone-Binding Proteins
D002352:Carrier Proteins; D004229:Dithiothreitol; D007457:Iodine Radioisotopes; D008565:Membrane Proteins; D009249:NADP; D012965:Sodium Chloride; D013963:Thyroid Hormones; D014284:Triiodothyronine; D000097810:Thyroid Hormone-Binding Proteins