[3H]BQ-123, a highly specific and reversible radioligand for the endothelin ETA receptor subtype

Eur J Pharmacol. 1995 Feb 14;274(1-3):1-6. doi: 10.1016/0014-2999(94)00670-3.

Abstract

The mode of binding of [3H]BQ-123 (cyclo(-D-Trp-D-Asp-[prolyl-3,4 (n)-[3H]]Pro-D-Val-Leu)), an endothelin receptor antagonist radioligand, was evaluated in the human neuroblastoma cell line SK-N-MC at 37 degrees C. Scatchard analysis indicated the presence of a single class of [3H]BQ-123 binding sites with a high affinity of 3.2 nM. [3H]BQ-123 binding achieved steady state within 7 min and dissociated with a half-time of 1.4 min, while [125I] endothelin-1 binding barely reached a steady state even after 6 h and showed little dissociation. [3H]BQ-123 binding was sensitive to endothelin-1 and endothelin-2 (Ki values = 0.058 and 0.10 nM, respectively) and the endothelin ETA receptor-selective antagonist BQ-123 (Ki = 3.3 nM), while showing low affinity for endothelin-3 (Ki = 50 nM), the endothelin ETB receptor-selective agonist BQ-3020 (Ki = 970 nM) and other bioactive peptides. Thus, [3H]BQ-123 is a specific and reversible radioligand for endothelin ETA receptors. The rapid reversibility of [3H]BQ-123 binding should provide a tool for estimating the equilibrium inhibition constants (Ki values) of various compounds for endothelin ETA receptors.

MeSH terms

  • Amino Acid Sequence
  • Azepines / metabolism
  • Binding, Competitive
  • Endothelin Receptor Antagonists*
  • Endothelins / antagonists & inhibitors
  • Endothelins / metabolism*
  • Humans
  • Indoles / metabolism
  • Isotope Labeling
  • Molecular Sequence Data
  • Neuroblastoma / metabolism
  • Neuroblastoma / pathology
  • Oligopeptides / metabolism
  • Peptides, Cyclic / metabolism*
  • Radioligand Assay
  • Receptor, Endothelin A
  • Receptors, Endothelin / metabolism
  • Structure-Activity Relationship
  • Tritium
  • Tumor Cells, Cultured

Substances

  • Azepines
  • Endothelin Receptor Antagonists
  • Endothelins
  • Indoles
  • Oligopeptides
  • Peptides, Cyclic
  • Receptor, Endothelin A
  • Receptors, Endothelin
  • Tritium
  • cyclo(sulfoalanyl-prolyl-valyl-leucyl-tryptophyl)
  • BQ 485
  • BQ 610
  • FR 139317
  • cyclo(Trp-Asp-Pro-Val-Leu)