title: Effect of protease inhibitors and substrates on 3,5,3'-triiodothyronine binding to rat liver nuclear receptors.
abstract: The effect of protease inhibitors N-tosyl-L-phenylalanine chloromethyl ketone (TPCK) and N-carbobenzoxy-L-phenylalanine chloromethyl ketone (ZPCK) at concentrations ranging from 1.5 x 10(-6) mol/l to 1.5 x 10(-4) mol/l on the specific binding of 3,5,3'-triiodothyronine (T3) to rat liver nuclear receptors was evaluated. Both TPCK at 1.5 x 10(-5) mol/l (P < 0.001) and ZPCK at 1.5 x 10(-6) mol/l (P < 0.05) showed the inhibition of T3 binding to nuclear receptors. The evaluation of T3 binding data following separation of unbound TPCK from treated nuclear receptors on a Sephadex G-25 column showed that the inhibition of T3 binding was irreversible, since a substantial decrease of the equilibrium association constant (Ka) was found when compared to the mock samples lacking TPCK. In addition, similar inhibitory effect on T3 specific binding to rat liver nuclear receptors was found at 5.0 x 10(-4) mol/l by protease substrates L-tyrosine benzyl ester (P < 0.001), L-tyrosine ethyl ester (P < 0.02), L-tryptophan methyl ester (P < 0.02) and L-tryptophan ethyl ester (P < 0.05). The data suggest that: 1. both protease inhibitors and substrates tested inhibited T3 specific binding to T3 nuclear receptors; 2. this may support the hypothesis that T3 receptors like other receptors encoded by c-erbA gene possess a site that recognize both serine protease inhibitors and substrates in its hormone binding domain.
authors.affiliation: Institute of Experimental Endocrinology, Slovak Academy of Sciences, Bratislava, CSFR.
authors.forename: J
authors.initials: J
authors.lastname: Brtko
authors.forename: J
authors.initials: J
authors.lastname: Knopp
authors.forename: P
authors.initials: P
authors.lastname: Filipcík
authors.forename: M E
authors.initials: ME
authors.lastname: Baker
mesh_terms: D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism*; D002850:Chromatography, Gel; D010450:Endopeptidases / Q000378:metabolism*; D004952:Esters / Q000494:pharmacology; D008099:Liver / Q000648:ultrastructure*; D008297:Male; D011480:Protease Inhibitors / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D011988:Receptors, Thyroid Hormone / Q000187:drug effects / Q000502:physiology*; D014108:Tosylphenylalanyl Chloromethyl Ketone / Q000494:pharmacology; D014284:Triiodothyronine / Q000378:metabolism*; D014364:Tryptophan / Q000031:analogs & derivatives / Q000494:pharmacology; D014443:Tyrosine / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: D000590:Amino Acid Chloromethyl Ketones; D004952:Esters; D011480:Protease Inhibitors; D011988:Receptors, Thyroid Hormone; D014284:Triiodothyronine; C006735:ZPCK; D014108:Tosylphenylalanyl Chloromethyl Ketone; D014443:Tyrosine; D014364:Tryptophan; D010450:Endopeptidases