D-Myo-inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] and D-myo-inositol 4,5-bisphosphate (10(-6) mol/L) displaced specifically bound L-[125I] T4 from human erythrocyte membranes in vitro by up to 80%. D-Myo-inositol 1,3,4,5-tetrakisphosphate, D-myo-inositol 1-monophosphate, and D-myo-inositol 1,4-bisphosphate were ineffective in decreasing thyroid hormone binding to membranes. The effect of Ins(1,4,5)P3 on high affinity binding reflected a change in Kd (5.8 x 10(-11) vs. 1.5 x 10(-11) mol/L) and binding capacity (15 vs. 2 fmol/mg membrane protein) in the absence and presence of Ins(1,4,5)P3, respectively. Ins(1,4,5)P3 also displaced T3 from red cell membranes. Thus, selected inositol phosphates regulate the abundance of sites available for binding of thyroid hormone by human red cell membranes. This stereospecific action of inositol phosphates is among several plasma membrane effects recently described for these members of the signal-transducing phosphoinositide pathway.