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ACS Cent Sci. 2018 Feb 28;4(2):166-179. doi: 10.1021/acscentsci.7b00237. Epub 2018 Jan 16.

Conditional and Reversible Activation of Class A and B G Protein-Coupled Receptors Using Tethered Pharmacology.

Author information

1
Department of Chemistry and Center for Integrated Protein Science, LMU Munich, 81377 Munich, Germany.
2
Institute of Metabolism and Systems Research (IMSR), University of Birmingham, B15 2TT, Birmingham, United Kingdom.
3
Centre for Endocrinology, Diabetes and Metabolism, Birmingham Health Partners, Birmingham, B15 2TH, United Kingdom, and COMPARE University of Birmingham and University of Nottingham Midlands.
4
Section of Cell Biology and Functional Genomics, Department of Medicine, Imperial College London, London, W12 0NN, United Kingdom.
5
Section of Investigative Medicine, Division of Diabetes, Endocrinology and Metabolism, Imperial College London, London, W12 0NN, United Kingdom.
6
Institut des Biomolécules Max Mousseron, UMR 5247 CNRS-Université Montpellier-ENSCM, Faculté de Pharmacie, 15 Avenue Charles Flahault, BP 14491, 34093 Montpellier Cedex 05, France.

Abstract

Understanding the activation and internalization of G protein-coupled receptors (GPCRs) using conditional approaches is paramount to developing new therapeutic strategies. Here, we describe the design, synthesis, and testing of ExONatide, a benzylguanine-linked peptide agonist of the glucagon-like peptide-1 receptor (GLP-1R), a class B GPCR required for maintenance of glucose levels in humans. ExONatide covalently binds to SNAP-tagged GLP-1R-expressing cells, leading to prolonged cAMP generation, Ca2+ rises, and intracellular retention of the receptor. These effects were readily switched OFF following cleavage of the introduced disulfide bridge using the cell-permeable reducing agent beta-mercaptoethanol (BME). A similar approach could be extended to a class A GPCR using GhrelON, a benzylguanine-linked peptide agonist of the growth hormone secretagogue receptor 1a (GHS-R1a), which is involved in food intake and growth. Thus, ExONatide and GhrelON allow SNAP-tag-directed activation of class A and B GPCRs involved in gut hormone signaling in a reversible manner. This tactic, termed reductively cleavable agONist (RECON), may be useful for understanding GLP-1R and GHS-R1a function both in vitro and in vivo, with applicability across GPCRs.

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