Different stability of ligand-receptor complex formed with two endothelin receptor species, ETA and ETB.
There are at least two types of endothelin receptors, ETA and ETB, present in various tissues. We found that although biotinylated ET-1 could bind to both ETA and ETB receptors, the stability of the formed ligand-receptor complexes was different. When the preformed complexes of receptor (solubilized from canine brain and lung membranes) and biotinylated ET-1 were subjected to avidin agarose column chromatography, most of the ETA activity was recovered in the pass-through fraction and the remainder was recovered in the 0.5 M KSCN eluate as ligand-free forms. On the other hand, the ETB activity bound firmly to the avidin agarose column was eluted with 1.5 M KSCN. The detection of the complex of 125I-ET-1 and its receptor by SDS-PAGE run at a low temperature was only possible with the ETB fractions and the complex of 125I-ET-1 and ETA was unstable during the separation. These results suggest that the conformation of the ligand binding sites of canine ETA and ETB as well as the stability of their ligand-receptor complexes to SDS are significantly different. Similar observations were also obtained for human ETA and ETB receptors.
Takasuka
T
T
Department of Molecular Genetics, Nippon Roche Research Center, Kanagawa.
Akiyama
N
N
Horii
I
I
Furuichi
Y
Y
Watanabe
T
T
D000818:Animals; D001665:Binding Sites; D001921:Brain / Q000378:metabolism; D002846:Chromatography, Affinity; D004285:Dogs; D016232:Endothelins / Q000378:metabolism*; D007700:Kinetics; D008168:Lung / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000145:classification / Q000302:isolation & purification / Q000378:metabolism*; D017466:Receptors, Endothelin
D016232:Endothelins; D011956:Receptors, Cell Surface; D017466:Receptors, Endothelin