We evaluated whether selected hormones, 3,5,3'-triiodothyronine (T3), hydrocortisone (HC) or insulin, would influence the binding (saturable, stereospecific, and of high affinity) of L-tryptophan to rat hepatic nuclei or nuclear envelopes. T3 (10(-14) to 10(-10) M) appreciably inhibited in vitro L-(5-3H) tryptophan binding to hepatic nuclei and T3 (10(-16) to 10(-4) M) appreciably ameliorated the inhibitory effect of unlabeled tryptophan (10(-4) M) on such binding. In vivo tryptophan administration (1 h) stimulated hepatic protein synthesis but the addition of T3 negated such stimulation. HC (10(-12) to 10(-4) M) did not affect and insulin (10(-16) to 10(-4) M) had only a small inhibitory effect on 3H-tryptophan binding to hepatic nuclei, but each (10(-12) to 10(-4) M) when added to unlabeled tryptophan (10(-4) M) diminished the inhibitory binding effect of unlabeled tryptophan alone. Thus, T3 competes with tryptophan for hepatic nuclear tryptophan binding and also negates tryptophan's stimulatory effect on hepatic protein synthesis.