Isolation of human cardiac endothelin receptors by a peptide-receptor mobility shift assay.
1. A peptide-protein mobility shift assay has been developed, using native polyacrylamide-gel electrophoresis, that enables the isolation of de-natured receptor proteins from small amounts of human cardiac tissue. 2. Radiolabelled endothelin-1 and related peptides were used to identify and isolate endothelin receptors from partially purified membrane extracts of human atrial tissue. 3. Binding analysis using radiolabelled endothelin-1 gave an equilibrium dissociation constant (Kd) of 2 nmol/l, similar to results from binding experiments conducted directly on tissue. 4. Peptide-receptor complexes were electroeluted from native gels and dissociated. Receptor material was characterized by dot-immunobinding analysis of eluates using an antibody raised against a predicted human endothelin receptor sequence.
Persidis
A
A
University of Cambridge Department of Medicine, U.K.
Harcombe
A A
AA
Davenport
A P
AP
Kuc
R E
RE
Plumpton
C
C
Weissberg
P L
PL
D001345:Autoradiography; D004591:Electrophoresis, Polyacrylamide Gel; D016232:Endothelins; D006325:Heart Atria / Q000737:chemistry; D006801:Humans; D009206:Myocardium / Q000737:chemistry*; D017466:Receptors, Endothelin / Q000302:isolation & purification*
D016232:Endothelins; D017466:Receptors, Endothelin