Identification of ETB receptor subtypes using linear and truncated analogs of ET.
Using canine spleen and lung membranes as model systems, we have shown the presence of subtypes of ETB receptors. This classification was done based on the binding profiles of various analogs of ET-1 that have been identified as ETB-selective. Saturation binding experiments performed with [125I] ET-3 and [125I] IRL-1260 (ETB-selective ligands) indicated that [125I] IRL-1620 labeled 80-90% of [125I] ET-3 binding sites in canine lung, whereas in canine spleen, the binding of [125I] IRL-1620 was 10-20% of [125I] ET-3 binding. In addition, competition binding experiments using ETB-selective agonists [Ala1,3,11,15]-ET-1 (also known as 4-Ala ET-1), N-acetyl-[Ala11,15] ET-1 (6-21) also known as BQ3020 and Suc-[Glu9, Ala11,15] ET-1 (8-21) also known as IRL-1620 indicated that all three ligands displaced [125I] ET-3 from canine lung membranes with approximately 500-1000 fold greater affinity than canine spleen membranes. On the other hand, ET-1, ET-3, S6a, S6b, and S6d displayed very similar IC50 values in both preparations, except S6c which was approximately 20 fold less potent in canine spleen compared to lung. These data indicate that ETB receptors present in canine spleen are different from those present in lung and that ETB-selective linear as well as truncated analogs of ET-1 are good tools to identify these subtypes of ETB receptors.
Nambi
P
P
Department of Renal Pharmacology, SmithKline Beecham Pharmaceuticals, King of Prussia, PA, USA.
Pullen
M
M
Brooks
D P
DP
Gellai
M
M
D000818:Animals; D001667:Binding, Competitive; D002462:Cell Membrane / Q000378:metabolism; D004285:Dogs; D019334:Endothelin-3 / Q000378:metabolism; D016232:Endothelins / Q000378:metabolism*; D007457:Iodine Radioisotopes; D008168:Lung / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism; D017466:Receptors, Endothelin / Q000145:classification / Q000378:metabolism*; D013154:Spleen / Q000378:metabolism
D019334:Endothelin-3; D016232:Endothelins; D007457:Iodine Radioisotopes; D010446:Peptide Fragments; D017466:Receptors, Endothelin; C074308:sovateltide