title: Tosyl-lysyl chloromethane alters glucocorticoid-receptor complex nuclear binding and physical properties.
abstract: We have investigated the effect of tosyl-lysyl chloromethane (a serine proteinase inhibitor) on rat liver cytosolic glucocorticoid-receptor complex binding to isolated nuclei. Tosyl-lysyl chloromethane (1-2 mM) significantly blocked nuclear binding when added before (but not after) thermal activation. Fifty percent inhibition occurred at about 1 mM tosyl-lysyl chloromethane. Interestingly, several other serine proteinase inhibitors (tosyl-phenylalanyl chloromethane, phenylmethyl sulphonylfluoride, and diisopropyl fluorophosphate) also depressed glucocorticoid-receptor complex nuclear binding. Tosyl-lysyl chloromethane, like molybdate, inhibited dilution-induced nuclear binding at low temperature, altered the DEAE-cellulose binding characteristics of heat-treated glucocorticoid-receptor complexes, and caused glucocorticoid-receptor complexes to sediment at about 9-10S (control complexes sedimented at 7-8S) in low salt-sucrose density gradients. Overall, these results suggest that tosyl-lysyl chloromethane modulates several properties of the glucocorticoid-receptor complex, that tosyl-lysyl chloromethane effects resemble those of molybdate, and that a serine proteinase(s) could be involved in the mechanism of glucocorticoid-receptor complex activation into a nuclear binding form.
mesh_terms: D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology*; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism*; D002499:Centrifugation, Density Gradient; D003080:Cold Temperature; D003600:Cytosol / Q000378:metabolism; D003636:DEAE-Cellulose / Q000378:metabolism; D004355:Drug Stability; D006358:Hot Temperature; D008099:Liver / Q000648:ultrastructure*; D008297:Male; D008982:Molybdenum / Q000494:pharmacology; D011480:Protease Inhibitors; D011506:Proteins; D051381:Rats; D011919:Rats, Inbred Strains; D011965:Receptors, Glucocorticoid / Q000187:drug effects / Q000378:metabolism*; D011987:Receptors, Steroid / Q000378:metabolism*; D013438:Sulfhydryl Compounds / Q000494:pharmacology; D014107:Tosyllysine Chloromethyl Ketone / Q000494:pharmacology*
chemical_list: D000590:Amino Acid Chloromethyl Ketones; D011480:Protease Inhibitors; D011506:Proteins; D011965:Receptors, Glucocorticoid; D011987:Receptors, Steroid; D013438:Sulfhydryl Compounds; D014107:Tosyllysine Chloromethyl Ketone; D008982:Molybdenum; D003636:DEAE-Cellulose