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
AU - Nasteska, Daniela
AU - Leippe, Philipp
AU - Gailer, Manuel
AU - Buenaventura, Teresa
AU - Kanda, Nisha
AU - Jones, Ben
AU - N'Kadmi, Celine
AU - Baneres, Jean-Louis
AU - Marie, Jacky
AU - Tomas, Alejandra
AU - Trauner, Dirk
AU - Hoffmann-Roder, Anja
AU - Hodson, David
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 a cell-permeable reducing agent. 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 a cell-permeable reducing agent. 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.
U2 - 10.1021/acscentsci.7b00237
DO - 10.1021/acscentsci.7b00237
M3 - Article
SN - 2374-7951
VL - 4
SP - 166
EP - 179
JO - ACS Central Science
JF - ACS Central Science
IS - 2
ER -