TY - JOUR
T1 - Conditional and Reversible Activation of Class A and B G Protein-Coupled Receptors Using Tethered Pharmacology
AU - Podewin, Tom
AU - Ast, Julia
AU - Broichhagen, Johannes
AU - Fine, Nicholas H.F.
AU - Nasteska, Daniela
AU - Leippe, Philipp
AU - Gailer, Manuel
AU - Buenaventura, Teresa
AU - Kanda, Nisha
AU - Jones, Ben J.
AU - M'Kadmi, Celine
AU - Baneres, Jean Louis
AU - Marie, Jacky
AU - Tomas, Alejandra
AU - Trauner, Dirk
AU - Hoffmann-Röder, Anja
AU - Hodson, David J.
N1 - Funding Information:
We are grateful to the SFB749 (T.P., M.G., and A.H.R.), the SFB TRR 152 (J.B. and D.T.), the SFB1032 (P.L. and D.T.), and the Center for Integrated Protein Science Munich (CIPSM) (T.P., J.B., P.L., M.G., D.T., and A.H.R.). T.P. was supported by an EASD Albert-Renold Young Scientist Fellowship (94571). A.T. was supported by an MRC Project Grant (MR/M012646/1) and a Diabetes UK Early-Career Small Grant (16/0005441). D.J.H. was supported by Diabetes UK. R.D. Lawrence (12/0004431), EFSD/Novo Nordisk Rising Star and Birmingham Fellowships, a Wellcome Trust Institutional Support Award, and an MRC Project Grant (MR/N00275X/1). This project has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (Starting Grant 715884 to D.J.H. and Advanced Grant 268795 to D.T.). MIN6B1 cells were provided by Prof. Philippe Halban (University of Geneva, Switzerland) with permission from Prof. Jun-ichi Miyazaki (University of Osaka) who produced the maternal MIN6 cell line.
Publisher Copyright:
© 2018 American Chemical Society.
Copyright:
Copyright 2018 Elsevier B.V., All rights reserved.
PY - 2018/2/28
Y1 - 2018/2/28
N2 - 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.
AB - 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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U2 - 10.1021/acscentsci.7b00237
DO - 10.1021/acscentsci.7b00237
M3 - Article
AN - SCOPUS:85042721825
VL - 4
SP - 166
EP - 179
JO - ACS Central Science
JF - ACS Central Science
SN - 2374-7943
IS - 2
ER -