Dynamics of GLP-1R peptide agonist engagement are correlated with kinetics of G protein activation

Giuseppe Deganutti; Yi-Lynn Liang; Xin Zhang; Maryam Khoshouei; Lachlan Clydesdale; Matthew J Belousoff; Hari Venugopal; Tin T Truong; Alisa Glukhova; Andrew N Keller; Karen J Gregory; Katie Leach; Arthur Christopoulos; Radostin Danev; Christopher A Reynolds; Peishen Zhao; Patrick M Sexton; Denise Wootten

doi:10.1038/s41467-021-27760-0

Dynamics of GLP-1R peptide agonist engagement are correlated with kinetics of G protein activation

Nat Commun. 2022 Jan 10;13(1):92. doi: 10.1038/s41467-021-27760-0.

Authors

Giuseppe Deganutti^#^{1

2}, Yi-Lynn Liang^#^{3

4}, Xin Zhang^#^{3

5}, Maryam Khoshouei^#^{6

7}, Lachlan Clydesdale^#³, Matthew J Belousoff^{3

5}, Hari Venugopal⁸, Tin T Truong³, Alisa Glukhova^{3

9}, Andrew N Keller³, Karen J Gregory³, Katie Leach³, Arthur Christopoulos^{3

5}, Radostin Danev¹⁰, Christopher A Reynolds^{11

12}, Peishen Zhao^{13

14}, Patrick M Sexton^{15

16}, Denise Wootten^{17

18}

Affiliations

¹ Centre for Sport, Exercise and Life Sciences, Coventry University, CV1 5FB, Coventry, UK.
² School of Biological Sciences, University of Essex, Colchester, CO4 3SQ, UK.
³ Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC, 3052, Australia.
⁴ Confo Therapeutics, Technologiepark 94, Ghent (Zwijnaarde), 9052, Belgium.
⁵ ARC Centre for Cryo-electron Microscopy of Membrane Proteins, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC, 3052, Australia.
⁶ Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, 82152, Martinsried, Germany.
⁷ Novartis Institutes for Biomedical Research, Novartis Pharma AG, Basel, Switzerland.
⁸ Ramaciotti Centre for Cryo-Electron Microscopy, Monash University, Clayton, VIC, 3168, Australia.
⁹ Walter and Eliza Hall Institute, Parkville, VIC, 3052, Australia.
¹⁰ Graduate School of Medicine, University of Tokyo, N415, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.
¹¹ Centre for Sport, Exercise and Life Sciences, Coventry University, CV1 5FB, Coventry, UK. [email protected].
¹² School of Biological Sciences, University of Essex, Colchester, CO4 3SQ, UK. [email protected].
¹³ Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC, 3052, Australia. [email protected].
¹⁴ ARC Centre for Cryo-electron Microscopy of Membrane Proteins, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC, 3052, Australia. [email protected].
¹⁵ Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC, 3052, Australia. [email protected].
¹⁶ ARC Centre for Cryo-electron Microscopy of Membrane Proteins, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC, 3052, Australia. [email protected].
¹⁷ Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC, 3052, Australia. [email protected].
¹⁸ ARC Centre for Cryo-electron Microscopy of Membrane Proteins, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC, 3052, Australia. [email protected].

^# Contributed equally.

Abstract

The glucagon-like peptide-1 receptor (GLP-1R) has broad physiological roles and is a validated target for treatment of metabolic disorders. Despite recent advances in GLP-1R structure elucidation, detailed mechanistic understanding of how different peptides generate profound differences in G protein-mediated signalling is still lacking. Here we combine cryo-electron microscopy, molecular dynamics simulations, receptor mutagenesis and pharmacological assays, to interrogate the mechanism and consequences of GLP-1R binding to four peptide agonists; glucagon-like peptide-1, oxyntomodulin, exendin-4 and exendin-P5. These data reveal that distinctions in peptide N-terminal interactions and dynamics with the GLP-1R transmembrane domain are reciprocally associated with differences in the allosteric coupling to G proteins. In particular, transient interactions with residues at the base of the binding cavity correlate with enhanced kinetics for G protein activation, providing a rationale for differences in G protein-mediated signalling efficacy from distinct agonists.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Allosteric Regulation
Baculoviridae / genetics
Baculoviridae / metabolism
Binding Sites
Cloning, Molecular
Cryoelectron Microscopy
Exenatide / chemistry*
Exenatide / genetics
Exenatide / metabolism
Gene Expression
Genetic Vectors / chemistry
Genetic Vectors / metabolism
Glucagon-Like Peptide 1 / chemistry*
Glucagon-Like Peptide 1 / genetics
Glucagon-Like Peptide 1 / metabolism
Glucagon-Like Peptide-1 Receptor / chemistry*
Glucagon-Like Peptide-1 Receptor / genetics
Glucagon-Like Peptide-1 Receptor / metabolism
HEK293 Cells
Humans
Kinetics
Ligands
Molecular Dynamics Simulation
Mutation
Oxyntomodulin / chemistry*
Oxyntomodulin / genetics
Oxyntomodulin / metabolism
Protein Binding
Protein Conformation, alpha-Helical
Protein Conformation, beta-Strand
Protein Interaction Domains and Motifs
Recombinant Proteins / chemistry
Recombinant Proteins / genetics
Recombinant Proteins / metabolism
Structure-Activity Relationship

Substances

GLP1R protein, human
Glucagon-Like Peptide-1 Receptor
Ligands
Oxyntomodulin
Recombinant Proteins
Glucagon-Like Peptide 1
Exenatide