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.2010:485:103-21.
doi: 10.1016/B978-0-12-381296-4.00006-3.

Ghrelin receptor: high constitutive activity and methods for developing inverse agonists

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Ghrelin receptor: high constitutive activity and methods for developing inverse agonists

Sylvia Els et al. Methods Enzymol.2010.

Abstract

The ghrelin receptor is a G protein-coupled receptor (GPCR) mainly distributed in the brain, and also expressed in peripheral tissues. Remarkably, the ghrelin receptor possesses a naturally high constitutive activity representing 50% of its maximal activity. Its endogenous ligand ghrelin is the only known orexigenic gastrointestinal peptide and plays a central role in the regulation of appetite, food intake, and energy homeostasis. Reducing the constitutive activity of the ghrelin receptor by inverse agonists is the strategy adopted by our group to develop anti-obesity drugs. Therefore, short peptides were synthesized and showed high inverse agonist potency toward the ghrelin receptor. This review describes the methods used to synthesize the peptides and to evaluate their biological activity. Peptide synthesis was performed on solid phase using a Fmoc/tBu-strategy. Peptide potency was measured with a signal transduction assay, the inositol trisphosphate turnover assay, adapted to a receptor expressing constitutive activity.

Copyright © 2010 Elsevier Inc. All rights reserved.

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