A Hydrogen-Bonded Polar Network in the Core of the Glucagon-Like Peptide-1 Receptor Is a Fulcrum for Biased Agonism: Lessons from Class B Crystal Structures

Denise Wootten, Christopher A Reynolds, Cassandra Koole, Kevin J Smith, Juan C Mobarec, John Simms, Tezz Quon, Thomas Coudrat, Sebastian G B Furness, Laurence J Miller, Arthur Christopoulos, Patrick M Sexton

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

The glucagon-like peptide 1 (GLP-1) receptor is a class B G protein-coupled receptor (GPCR) that is a key target for treatments for type II diabetes and obesity. This receptor, like other class B GPCRs, displays biased agonism, though the physiologic significance of this is yet to be elucidated. Previous work has implicated R2.60(190), N3.43(240), Q7.49(394), and H6.52(363) as key residues involved in peptide-mediated biased agonism, with R2.60(190), N3.43(240), and Q7.49(394) predicted to form a polar interaction network. In this study, we used novel insight gained from recent crystal structures of the transmembrane domains of the glucagon and corticotropin releasing factor 1 (CRF1) receptors to develop improved models of the GLP-1 receptor that predict additional key molecular interactions with these amino acids. We have introduced E6.53(364)A, N3.43(240)Q, Q7.49(394)N, and N3.43(240)Q/Q7.49(394)N mutations to probe the role of predicted H-bonding and charge-charge interactions in driving cAMP, calcium, or extracellular signal-regulated kinase (ERK) signaling. A polar interaction between E6.53(364) and R2.60(190) was predicted to be important for GLP-1- and exendin-4-, but not oxyntomodulin-mediated cAMP formation and also ERK1/2 phosphorylation. In contrast, Q7.49(394), but not R2.60(190)/E6.53(364) was critical for calcium mobilization for all three peptides. Mutation of N3.43(240) and Q7.49(394) had differential effects on individual peptides, providing evidence for molecular differences in activation transition. Collectively, this work expands our understanding of peptide-mediated signaling from the GLP-1 receptor and the key role that the central polar network plays in these events.

Original languageEnglish
Pages (from-to)335-47
Number of pages13
JournalMolecular Pharmacology
Volume89
Issue number3
Early online date31 Jan 2016
DOIs
Publication statusPublished - Mar 2016
Externally publishedYes

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Hydrogen
Peptides
Oxyntomodulin
Calcium
Corticotropin-Releasing Hormone Receptors
Mutation
Glucagon-Like Peptide 1
Extracellular Signal-Regulated MAP Kinases
G-Protein-Coupled Receptors
Type 2 Diabetes Mellitus
Obesity
Phosphorylation
Amino Acids
Glucagon-Like Peptide-1 Receptor

Keywords

  • Animals
  • CHO Cells
  • Cricetinae
  • Cricetulus
  • Crystallization
  • Glucagon-Like Peptide 1
  • Humans
  • Hydrogen Bonding
  • Models, Molecular
  • Peptide Fragments
  • Protein Binding
  • Protein Structure, Secondary
  • Journal Article
  • Research Support, Non-U.S. Gov't

Cite this

A Hydrogen-Bonded Polar Network in the Core of the Glucagon-Like Peptide-1 Receptor Is a Fulcrum for Biased Agonism : Lessons from Class B Crystal Structures. / Wootten, Denise; Reynolds, Christopher A; Koole, Cassandra; Smith, Kevin J; Mobarec, Juan C; Simms, John; Quon, Tezz; Coudrat, Thomas; Furness, Sebastian G B; Miller, Laurence J; Christopoulos, Arthur; Sexton, Patrick M.

In: Molecular Pharmacology, Vol. 89, No. 3, 03.2016, p. 335-47.

Research output: Contribution to journalArticle

Wootten, D, Reynolds, CA, Koole, C, Smith, KJ, Mobarec, JC, Simms, J, Quon, T, Coudrat, T, Furness, SGB, Miller, LJ, Christopoulos, A & Sexton, PM 2016, 'A Hydrogen-Bonded Polar Network in the Core of the Glucagon-Like Peptide-1 Receptor Is a Fulcrum for Biased Agonism: Lessons from Class B Crystal Structures' Molecular Pharmacology, vol. 89, no. 3, pp. 335-47. https://doi.org/10.1124/mol.115.101246
Wootten, Denise ; Reynolds, Christopher A ; Koole, Cassandra ; Smith, Kevin J ; Mobarec, Juan C ; Simms, John ; Quon, Tezz ; Coudrat, Thomas ; Furness, Sebastian G B ; Miller, Laurence J ; Christopoulos, Arthur ; Sexton, Patrick M. / A Hydrogen-Bonded Polar Network in the Core of the Glucagon-Like Peptide-1 Receptor Is a Fulcrum for Biased Agonism : Lessons from Class B Crystal Structures. In: Molecular Pharmacology. 2016 ; Vol. 89, No. 3. pp. 335-47.
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AU - Smith, Kevin J

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AU - Quon, Tezz

AU - Coudrat, Thomas

AU - Furness, Sebastian G B

AU - Miller, Laurence J

AU - Christopoulos, Arthur

AU - Sexton, Patrick M

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KW - CHO Cells

KW - Cricetinae

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KW - Crystallization

KW - Glucagon-Like Peptide 1

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KW - Models, Molecular

KW - Peptide Fragments

KW - Protein Binding

KW - Protein Structure, Secondary

KW - Journal Article

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

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