title: Characterization of [125I]endothelin-1 binding sites in rat cardiac membrane fragments.
abstract: Standard binding and displacement techniques were used to identify high-affinity binding sites for [125I]-labeled endothelin-1 (ET-1) in membranes harvested from the hearts of adult female Sprague-Dawley rats. A single population of binding sites was identified, with a KD of 0.20 +/- 0.03 nM at 37 degrees C, and a Bmax of 93.5 +/- 6.4 fmol/mg protein. Bound [125I]ET-1 was displaced by ET-1 (10(-13)-10(-8) M), with a Ki of 0.08 nM. Neither (-)Bay K 8644 (10(-11)-10(-5) M), prenylamine (10(-11)-10(-5) M), (+)-cis-diltiazem (10(-10)-10(-5) M), (-)D888 (10(-10)-10(-5) M), nicardipine (10(-10)-10(-5) M), lidoflazine (10(-11)-10(-5) M), flunarizine (10(-11)-10(-5) M), omega-conotoxin (10(-13)-10(-7) M), nor prazosin (10(-10)-10(-5) M) displaced the bound ligand. Binding occurred in the absence of Ca2+ and was absent in heat-denatured membranes. These results are interpreted to mean that [125I]ET-1 binds to a single class of high-affinity binding sites that differ from those occupied by known regulators of voltage activated L- and N-type Ca2+ channels.
authors: Department of Medicine, University of Melbourne, Victoria, Australia.
authors: X H
authors: XH
authors: Gu
authors: D J
authors: DJ
authors: Casley
authors: W G
authors: WG
authors: Nayler
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive / Q000187:drug effects; D016232:Endothelins; D066298:In Vitro Techniques; D007457:Iodine Radioisotopes; D008297:Male; D008969:Molecular Sequence Data; D009206:Myocardium / Q000378:metabolism*; D010455:Peptides / Q000032:analysis / Q000378:metabolism* / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000378:metabolism*; D017466:Receptors, Endothelin; D013552:Swine
chemical_list: D016232:Endothelins; D007457:Iodine Radioisotopes; D010455:Peptides; D011956:Receptors, Cell Surface; D017466:Receptors, Endothelin