Structural and functional diversity among agonist-bound states of the GLP-1 receptor

Cary, Brian P.; Deganutti, Giuseppe; Zhao, Peishen; Truong, Tin T.; Piper, Sarah; Liu, Xinyu; Belousoff, Matthew J.; Danev, Radostin; Sexton, Patrick M.; Wootten, Denise; Gellman, Samuel H.

doi:10.1038/s41589-021-00945-w

Cited by 27 publications

(90 citation statements)

References 72 publications

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“…Supporting both observations is the fact that the positive functional modulation () of LY298 and VU154 in Gi1 signalling assays is driven by positive binding modulation (), as the efficacy modulation () of the PAMs was negative to neutral (Figure 2J, S1E). These observations were consistent with recent studies that suggest the conformational dynamics between agonist and receptor were important for functional signalling (Bumbak et al, 2020;Cary et al, 2022;Deganutti et al, 2022;O'Connor et al, 2015).…”

Section: Discussionsupporting

confidence: 93%

Pharmacological hallmarks of allostery at the M4 muscarinic receptor elucidated through structure and dynamics

Vuckovic

Wang

Pham

et al. 2022

Preprint

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Allosteric modulation of G protein-coupled receptors (GPCRs) is a major paradigm in drug discovery. Despite decades of research, a molecular level understanding of the general principals that govern the myriad pharmacological effects exerted by GPCR allosteric modulators remains limited. The M4 muscarinic acetylcholine receptor (M4 mAChR) is a well-validated and clinically relevant allosteric drug target for several major psychiatric and cognitive disorders. Here, we present high-resolution cryo-electron microscopy structures of the M4 mAChR bound to a cognate Gi1 protein and the high affinity agonist, iperoxo, in the absence and presence of two different positive allosteric modulators, LY2033298 or VU0467154. We have also determined the structure of the M4 mAChR-Gi1 complex bound to its endogenous agonist, acetylcholine (ACh). Structural comparisons, together with molecular dynamics, mutagenesis, and pharmacological validations, have provided in-depth insights into the role of structure and dynamics in orthosteric and allosteric ligand binding, global mechanisms of receptor activation, cooperativity, probe-dependence, and species variability; all key hallmarks underpinning contemporary GPCR drug discovery.

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Section: Discussionsupporting

confidence: 93%

Pharmacological hallmarks of allostery at the M4 muscarinic receptor elucidated through structure and dynamics

Vuckovic

Wang

Pham

et al. 2022

Preprint

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“…While the ECD, where the peptide C-terminus initially recognizes class B1 GPCRs, remains less well-resolved than other regions, the resolution of structures determined through cryo-EM has drastically improved over the past five years through advances in sample preparation, data collection, and data processing, and is predicted to continue improving with sub-2.5 Å cryo-EM structure determination being increasingly routine for class B1 GPCR–G protein complexes [ 288 ]. Moreover, direct information on 3D conformational dynamics captured during vitrification can increasingly be extracted through sophisticated analytical methods [ 304 , 345 ] and such dynamics play critical roles in peptide recognition and modes of receptor activation that may not be apparent from high-resolution consensus reconstructions [ 304 , 332 , 346 , 347 ]. This information can be integrated with other methods for interrogating GPCR dynamics, including hydrogen deuterium exchange mass spectrometry (HDX-MS) [ 348 ], single-molecule FRET studies [ 349 ] and electron paramagnetic resonance (EPR) [ 350 , 351 ] and holds promise for further enhancing peptide drug design [ 348 , 352 ].…”

Section: Discussionmentioning

confidence: 99%

Targeting VIP and PACAP Receptor Signaling: New Insights into Designing Drugs for the PACAP Subfamily of Receptors

Piper

Zhao

et al. 2022

IJMS

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Pituitary Adenylate Cyclase-Activating Peptide (PACAP) and Vasoactive Intestinal Peptide (VIP) are neuropeptides involved in a diverse array of physiological and pathological processes through activating the PACAP subfamily of class B1 G protein-coupled receptors (GPCRs): VIP receptor 1 (VPAC1R), VIP receptor 2 (VPAC2R), and PACAP type I receptor (PAC1R). VIP and PACAP share nearly 70% amino acid sequence identity, while their receptors PAC1R, VPAC1R, and VPAC2R share 60% homology in the transmembrane regions of the receptor. PACAP binds with high affinity to all three receptors, while VIP binds with high affinity to VPAC1R and VPAC2R, and has a thousand-fold lower affinity for PAC1R compared to PACAP. Due to the wide distribution of VIP and PACAP receptors in the body, potential therapeutic applications of drugs targeting these receptors, as well as expected undesired side effects, are numerous. Designing selective therapeutics targeting these receptors remains challenging due to their structural similarities. This review discusses recent discoveries on the molecular mechanisms involved in the selectivity and signaling of the PACAP subfamily of receptors, and future considerations for therapeutic targeting.

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“…Previous data suggested that plasticity of the peptide-TMD engagement is relevant for activity at the glucagon-like peptide-1 receptor (GLP-1R) (Cary et al, 2022;Deganutti et al, 2022), a related class B1 GPCR, where the N-terminus of some peptides become partially or fully unbound in 3DVA associated with outward movement of ECL3. This dynamic behaviour was commonly observed in peptides that have residues that can destabilise helical secondary structure of the activation domain of those peptides (Johnson et al, 2021;Zhang et al, 2021;Cary et al, 2022). In contrast, all PTH1R-peptide complexes in the current study exhibited stable interactions between the peptide N-terminus and the receptor core (Video S1), suggesting that this interaction might be generally more stable than for agonists of the GLP-1R.…”

Section: Discussionmentioning

confidence: 99%

“…Full-length peptide agonists of the PTH1R bind in a two-step process (Castro et al, 2005), corresponding to sequential binding to the ECD and the TMD. Previous data suggested that plasticity of the peptide-TMD engagement is relevant for activity at the glucagon-like peptide-1 receptor (GLP-1R) (Cary et al, 2022;Deganutti et al, 2022), a related class B1 GPCR, where the N-terminus of some peptides become partially or fully unbound in 3DVA associated with outward movement of ECL3. This dynamic behaviour was commonly observed in peptides that have residues that can destabilise helical secondary structure of the activation domain of those peptides (Johnson et al, 2021;Zhang et al, 2021;Cary et al, 2022).…”

Section: Discussionmentioning

confidence: 99%

Molecular insights into peptide agonist engagement with the PTH1 receptor

Cary

Gerrard

Belousoff

et al. 2022

Preprint

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The parathyroid hormone (PTH) 1 receptor (PTH1R) is a class B1 G protein-coupled receptor (GPCR) that critically regulates skeletal development and calcium homeostasis. Despite extensive study, the molecular underpinnings of PTH1R stimulation by its cognate hormones, as well as by therapeutic agents, remain unclear. Here, we describe cryo-EM structures of the PTH1R in complex with active fragments of the two hormones, PTH and parathyroid hormone related protein (PTHrP), the peptidic drug abaloparatide, as well as the engineered tool compounds, long-acting PTH (LA-PTH) and the truncated peptide, M-PTH(1-14). We found that the N-terminus of each agonist that is critical for activity, engages the transmembrane bundle in a topologically similar fashion, which reflects similarities in measures of Gαs activation. The full-length peptides bind the extracellular domain (ECD) using a shared interface but induce subtly different ECD orientations relative to the transmembrane domain (TMD). In the structure bound to M-PTH, an agonist which only binds the TMD, the ECD is completely unresolved, demonstrating that the ECD is highly dynamic when unconstrained by a peptide. High resolutions enabled identification of water molecules near the peptide and G protein binding sites, some of which are structurally conserved with other class B1 GPCRs. Our results shed light on the action of orthosteric agonists of the PTH1R and provide a foundation for structure based-drug design.

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Structural and functional diversity among agonist-bound states of the GLP-1 receptor

Abstract: This document is the author's post-print version, incorporating any revisions agreed during the peer-review process. Some differences between the published version and this version may remain and you are advised to consult the published version if you wish to cite from it.

Cited by 27 publications

References 72 publications

Pharmacological hallmarks of allostery at the M4 muscarinic receptor elucidated through structure and dynamics

Pharmacological hallmarks of allostery at the M4 muscarinic receptor elucidated through structure and dynamics

Targeting VIP and PACAP Receptor Signaling: New Insights into Designing Drugs for the PACAP Subfamily of Receptors

Molecular insights into peptide agonist engagement with the PTH1 receptor

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