Dimerization of the human receptors for prostacyclin and thromboxane facilitates thromboxane receptor-mediated cAMP generation.
Prostacyclin (PGI(2)) and thromboxane (TxA(2)) are biological opposites; PGI(2), a vasodilator and inhibitor of platelet aggregation, limits the deleterious actions of TxA(2), a vasoconstrictor and platelet activator. The molecular mechanisms involved in the counterregulation of PGI(2)/TxA(2) signaling are unclear. We examined the interaction of the receptors for PGI(2) (IP) and TxA(2) (TPalpha). IP-induced cAMP and TP-induced inositol phosphate generation were unaltered when the receptors were co-expressed in HEK 293 cells (IP/TPalpha-HEK). TP-cAMP generation, in response to TP agonists or a TP-dependent isoprostane, iPE(2)III, was evident in IP/TPalpha-HEK and in aortic smooth muscle cells, but not in cells expressing either receptor alone, or in IP-deficient aortic smooth muscle cells. Augmentation of TP-induced cAMP generation, with the IP agonist cicaprost, was ablated in IP-deficient cells and was independent of direct IP signaling. IP/TPalpha heterodimers were formed constitutively when the receptors were co-expressed, with no overt changes in ligand binding to the individual receptor sites. However, despite inefficient binding of iPE(2)III to either the IP or TPalpha, expressed alone or in combination, robust cAMP generation was evident in IP/TPalpha-HEK, suggesting the formation of an alternative receptor site. Thus, IP/TPalpha dimerization was coincident with TP-cAMP generation, promoting a "PGI(2)-like" cellular response to TP activation. This represents a previously unknown mechanism by which IP may limit the cellular effects of TP.
Center for Experimental Therapeutics, University of Pennsylvania, 421 Curie Blvd., Philadelphia, PA 19104, USA.
Stephen J
SJ
Wilson
Aoife M
AM
Roche
Ekaterina
E
Kostetskaia
Emer M
EM
Smyth
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000378:metabolism; D000818:Animals; D001011:Aorta / Q000473:pathology; D001665:Binding Sites; D015153:Blotting, Western; D019086:Bridged Bicyclo Compounds, Heterocyclic; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors; D019281:Dimerization; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000737:chemistry; D011464:Epoprostenol / Q000031:analogs & derivatives* / Q000494:pharmacology; D005231:Fatty Acids, Unsaturated; D006801:Humans; D006834:Hydrazines / Q000494:pharmacology; D015151:Immunoblotting; D047468:Immunoprecipitation; D007295:Inositol Phosphates / Q000737:chemistry; D028421:Isoprostanes / Q000378:metabolism; D008024:Ligands; D051379:Mice; D009130:Muscle, Smooth / Q000166:cytology; D010974:Platelet Aggregation; D011485:Protein Binding; D011863:Radioimmunoassay; D044006:Receptors, Epoprostenol / Q000737:chemistry*; D017482:Receptors, Thromboxane / Q000737:chemistry*; D015398:Signal Transduction; D013997:Time Factors; D014162:Transfection
D019086:Bridged Bicyclo Compounds, Heterocyclic; D000939:Epitopes; D005231:Fatty Acids, Unsaturated; D006834:Hydrazines; D007295:Inositol Phosphates; D028421:Isoprostanes; D008024:Ligands; D044006:Receptors, Epoprostenol; D017482:Receptors, Thromboxane; D015124:8-Bromo Cyclic Adenosine Monophosphate; C045749:SQ 29548; D011464:Epoprostenol; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; C043867:cicaprost