Characterization of the prostanoid TP receptor population in human nonpregnant myometrium.
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.
Senchyna
M
M
Department of Obstetrics and Gynecology, McMaster University, Hamilton, Ontario, Canada.
Crankshaw
D J
DJ
D019796:15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; D000328:Adult; D001667:Binding, Competitive; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005231:Fatty Acids, Unsaturated / Q000378:metabolism; D005260:Female; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D008875:Middle Aged; D009215:Myometrium / Q000737:chemistry*; D011450:Prostaglandin Endoperoxides, Synthetic / Q000494:pharmacology; D017482:Receptors, Thromboxane / Q000032:analysis*; D013928:Thromboxane A2 / Q000031:analogs & derivatives / Q000494:pharmacology
D019086:Bridged Bicyclo Compounds, Heterocyclic; D005231:Fatty Acids, Unsaturated; D011450:Prostaglandin Endoperoxides, Synthetic; D017482:Receptors, Thromboxane; C061750:7-(3-(3-hydroxy-4-(4'-iodophenoxy)-1-butenyl)-7-oxabicyclo(2.2.1)heptan-2-yl)-5-heptenoic acid; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D013928:Thromboxane A2; D019796:15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid