In vitro effects of thyroid hormones on red blood cell Ca++-dependent ATPase activity

Thyroid hormones (TH) have been shown to exert a direct stimulatory effect on the Ca++-dependent ATPase from human and other mammalian erythrocytes. In this in vitro system, T4 has been shown to be more effective than T3. In the present study, TH effects on Ca++-dependent ATPase were investigated, using rabbit and human erythrocyte membranes, after preincubation with 10(-10) M T4, in the presence or in the absence of exogenous calmodulin (CaM) (5.10(-12) M to 5.10(-9) M). Ca++-dependent ATPase activity was measured as inorganic phosphate (Pi) release from 1 mM ATP. The results showed that basal Ca++-dependent ATPase activity in rabbits was moderately increased by T4 (1.44 +/- 0.05 vs 1.32 +/- 0.04 mumol Pi/mg protein/90 min, mean +/- SE; p less than 0.05). The time course of Pi release did not show any stimulatory effect of T4 during the first hour of incubation. The effect of T4 became apparent, however, 1 h after the addition of ATP (delta T4: 15%). With human membranes, T4 induced a relative stimulation of the Ca++-dependent ATPase of 8-10% (p less than 0.05) in experimental conditions where the enzyme was not maximally stimulated by CaM (delta CaM over basal activity: 5-40%). In conditions of high CaM stimulation (delta CaM: 50-320%), T4 had no effect. These results confirm that Ca++-dependent ATPase activity is increased by T4. The effect of T4 is small, and appears as a late event during incubation with ATP. Stimulation by T4 is expressed in states of low enzyme activation by CaM.(ABSTRACT TRUNCATED AT 250 WORDS)