We have used both functional and binding studies to fully characterize the prostanoid TP receptor in the myometrium from nonpregnant human donors. Both U-46,619 and I-BOP produced concentration-dependent contraction of human myometrial strips in vitro (pEC50 = 6.9 +/- 0.6; and 7.8 +/- 0.5, respectively). U-46,619-induced contractions were attenuated by the TP receptor antagonists: ICI 192,605 (pKB = 9.2 +/- 0.3); ICI D1,542 (pKB = 9.1 +/- 0.3); L670,596 (pKB = 8.6 +/- 0.3); GR 32,191 (pKB = 8.6 +/- 0.2); SQ 29,548 (pKB = 8.2 +/- 0.5); ONO 3,708 (pKB = 8.1 +/- 0.3) and BM 13,505 (pKB = 7.4 +/- 0.2). The binding of [125I]-BOP to human myometrial membranes was saturable, selective and displaceable. Equilibrium binding of [125I]-BOP identified one class of sites, Kd = 3.4 nM (pKd = 8.7 +/- 0.4) and a maximum binding of 323.1 +/- 361.5 fmol/mg protein. The addition of the nonhydrolyzable GTP analog GTP gamma S (100 microM) to the assay had no effect on [125I]-BOP binding. The Kd determined kinetically was 4.1 +/- 0.2 nM. TP receptor antagonists competed for [125I]-BOP binding: ICI D1,542 (pIC50 = 8.3 +/- 0.4); L670,596 (pIC50 = 7.9 +/- 0.1); ICI 192,605 (pIC50 = 7.2 +/- 0.1); ONO 3,708 (pIC50 = 7.2 +/- 0.04); SQ 29,548 (pIC50 = 7.2 +/- 0.1); GR 32,191 (pIC50 = 7.0 +/- 0.2); BM 13,505 (pIC50 = 6.8 +/- 0.1). The rank order of potency for the seven TP receptor antagonists in displacing [125I]-BOP from its binding site was correlated (r = 0.75) with the rank order of potency in inhibiting U-46,619-induced contraction of myometrial strips. Ligands selective for other prostanoid receptors were unable to significantly displace [125I]-BOP binding. These results are consistent with the notion that the human myometrial TP receptor is pharmacologically similar to the low affinity TP receptor in human platelets.