Structural Insight into Antibody-mediated Antagonism of the Glucagon-like Peptide-1 Receptor

Overview

Journal Sci Rep

Specialty Science

Date 2016 May 20

PMID 27196125

Citations 17

Authors

Stephanie Hennen

Janos T Kodra

Vladyslav Soroka

Berit O Krogh

Xiaoai Wu

Peter Kaastrup

Cathrine Orskov

Sif G Ronn

Gerd Schluckebier

Silvia Barbateskovic

Prafull S Gandhi

Steffen Reedtz-Runge

Affiliations

Soon will be listed here.

Abstract

The Glucagon-like peptide-1 receptor (GLP-1R) is a member of the class B G protein-coupled receptor (GPCR) family and a well-established target for the treatment of type 2 diabetes. The N-terminal extracellular domain (ECD) of GLP-1R is important for GLP-1 binding and the crystal structure of the GLP-1/ECD complex was reported previously. The first structure of a class B GPCR transmembrane (TM) domain was solved recently, but the full length receptor structure is still not well understood. Here we describe the molecular details of antibody-mediated antagonism of the GLP-1R using both in vitro pharmacology and x-ray crystallography. We showed that the antibody Fab fragment (Fab 3F52) blocked the GLP-1 binding site of the ECD directly and thereby acts as a competitive antagonist of native GLP-1. Interestingly, Fab 3F52 also blocked a short peptide agonist believed to engage primarily the transmembrane and extracellular loop region of GLP-1R, whereas functionality of an allosteric small-molecule agonist was not inhibited. This study has implications for the structural understanding of the GLP-1R and related class B GPCRs, which is important for the development of new and improved therapeutics targeting these receptors.

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References

Tibaduiza E, Chen C, Beinborn M . A small molecule ligand of the glucagon-like peptide 1 receptor targets its amino-terminal hormone binding domain. J Biol Chem. 2001; 276(41):37787-93. DOI: 10.1074/jbc.M106692200. View

Durocher Y, Perret S, Kamen A . High-level and high-throughput recombinant protein production by transient transfection of suspension-growing human 293-EBNA1 cells. Nucleic Acids Res. 2002; 30(2):E9. PMC: 99848. DOI: 10.1093/nar/30.2.e9. View

Lopez de Maturana R, Donnelly D . The glucagon-like peptide-1 receptor binding site for the N-terminus of GLP-1 requires polarity at Asp198 rather than negative charge. FEBS Lett. 2002; 530(1-3):244-8. DOI: 10.1016/s0014-5793(02)03492-0. View

Runge S, Wulff B, Madsen K, Brauner-Osborne H, Knudsen L . Different domains of the glucagon and glucagon-like peptide-1 receptors provide the critical determinants of ligand selectivity. Br J Pharmacol. 2003; 138(5):787-94. PMC: 1573731. DOI: 10.1038/sj.bjp.0705120. View

Thorens B . Expression cloning of the pancreatic beta cell receptor for the gluco-incretin hormone glucagon-like peptide 1. Proc Natl Acad Sci U S A. 1992; 89(18):8641-5. PMC: 49976. DOI: 10.1073/pnas.89.18.8641. View

Grace C, Perrin M, DiGruccio M, Miller C, Rivier J, Vale W . NMR structure and peptide hormone binding site of the first extracellular domain of a type B1 G protein-coupled receptor. Proc Natl Acad Sci U S A. 2004; 101(35):12836-41. PMC: 516482. DOI: 10.1073/pnas.0404702101. View

Emsley P, Cowtan K . Coot: model-building tools for molecular graphics. Acta Crystallogr D Biol Crystallogr. 2004; 60(Pt 12 Pt 1):2126-32. DOI: 10.1107/S0907444904019158. View

Hoare S . Mechanisms of peptide and nonpeptide ligand binding to Class B G-protein-coupled receptors. Drug Discov Today. 2005; 10(6):417-27. DOI: 10.1016/S1359-6446(05)03370-2. View

Pham V, Dong M, Wade J, Miller L, Morton C, Ng H . Insights into interactions between the alpha-helical region of the salmon calcitonin antagonists and the human calcitonin receptor using photoaffinity labeling. J Biol Chem. 2005; 280(31):28610-22. DOI: 10.1074/jbc.M503272200. View

10.

Knudsen L, Kiel D, Teng M, Behrens C, Bhumralkar D, Kodra J . Small-molecule agonists for the glucagon-like peptide 1 receptor. Proc Natl Acad Sci U S A. 2007; 104(3):937-42. PMC: 1783418. DOI: 10.1073/pnas.0605701104. View

11.

Runge S, Schimmer S, Oschmann J, Bruun Schiodt C, Knudsen S, Jeppesen C . Differential structural properties of GLP-1 and exendin-4 determine their relative affinity for the GLP-1 receptor N-terminal extracellular domain. Biochemistry. 2007; 46(19):5830-40. DOI: 10.1021/bi062309m. View

12.

Parthier C, Kleinschmidt M, Neumann P, Rudolph R, Manhart S, Schlenzig D . Crystal structure of the incretin-bound extracellular domain of a G protein-coupled receptor. Proc Natl Acad Sci U S A. 2007; 104(35):13942-7. PMC: 1955799. DOI: 10.1073/pnas.0706404104. View

13.

Runge S, Thogersen H, Madsen K, Lau J, Rudolph R . Crystal structure of the ligand-bound glucagon-like peptide-1 receptor extracellular domain. J Biol Chem. 2008; 283(17):11340-7. DOI: 10.1074/jbc.M708740200. View

14.

Pioszak A, Xu H . Molecular recognition of parathyroid hormone by its G protein-coupled receptor. Proc Natl Acad Sci U S A. 2008; 105(13):5034-9. PMC: 2278174. DOI: 10.1073/pnas.0801027105. View

15.

Pioszak A, Parker N, Suino-Powell K, Xu H . Molecular recognition of corticotropin-releasing factor by its G-protein-coupled receptor CRFR1. J Biol Chem. 2008; 283(47):32900-12. PMC: 2583312. DOI: 10.1074/jbc.M805749200. View

16.

Mapelli C, Natarajan S, Meyer J, Bastos M, Bernatowicz M, Lee V . Eleven amino acid glucagon-like peptide-1 receptor agonists with antidiabetic activity. J Med Chem. 2009; 52(23):7788-99. DOI: 10.1021/jm900752a. View

17.

Underwood C, Garibay P, Knudsen L, Hastrup S, Peters G, Rudolph R . Crystal structure of glucagon-like peptide-1 in complex with the extracellular domain of the glucagon-like peptide-1 receptor. J Biol Chem. 2009; 285(1):723-30. PMC: 2804221. DOI: 10.1074/jbc.M109.033829. View

18.

Adams P, Afonine P, Bunkoczi G, Chen V, Davis I, Echols N . PHENIX: a comprehensive Python-based system for macromolecular structure solution. Acta Crystallogr D Biol Crystallogr. 2010; 66(Pt 2):213-21. PMC: 2815670. DOI: 10.1107/S0907444909052925. View

19.

Patterson J, Ottaway N, Gelfanov V, Smiley D, Perez-Tilve D, Pfluger P . A novel human-based receptor antagonist of sustained action reveals body weight control by endogenous GLP-1. ACS Chem Biol. 2010; 6(2):135-45. DOI: 10.1021/cb1002015. View

20.

Caporale A, Sturlese M, Gesiot L, Zanta F, Wittelsberger A, Cabrele C . Side chain cyclization based on serine residues: synthesis, structure, and activity of a novel cyclic analogue of the parathyroid hormone fragment 1-11. J Med Chem. 2010; 53(22):8072-9. DOI: 10.1021/jm1008264. View