Fluorescence Correlation Spectroscopy in Drug Discovery: Study of Alexa532-endothelin 1 Binding to the Endothelin ETA Receptor to Describe the Pharmacological Profile of Natural Products

Overview

Journal ScientificWorldJournal

Publisher Wiley

Specialty Biology

Date 2012 May 25

PMID 22623909

Citations 2

Authors

Catherina Caballero-George

Thomas Sorkalla

Daniel Jakobs

Jessica Bolanos

Huzefa Raja

Carol Shearer

Eldredge Bermingham

Hanns Haberlein

Affiliations

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Abstract

Fluorescence correlation spectroscopy and the newly synthesized Alexa532-ET1 were used to study the dynamics of the endothelin ET(A) receptor-ligand complex alone and under the influence of a semisynthetic selective antagonist and a fungal extract on living A10 cells. Dose-dependent increase of inositol phosphate production was seen for Alexa532-ET1, and its binding was reduced to 8% by the selective endothelin ET(A) antagonist BQ-123, confirming the specific binding of Alexa532-ET1 to the endothelin ET(A) receptor. Two different lateral mobilities of the receptor-ligand complexes within the cell membrane were found allowing the discrimination of different states for this complex. BQ-123 showed a strong binding affinity to the "inactive" receptor state characterized by the slow diffusion time constant. A similar effect was observed for the fungal extract, which completely displaced Alexa532-ET1 from its binding to the "inactive" receptor state. These findings suggest that both BQ-123 and the fungal extract act as inverse agonists.

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