Glucagon-like peptide-1 receptor internalisation controls spatiotemporal signalling mediated by biased agonists

Fletcher, Madeleine; Halls, Michelle L.; Zhao, Peishen; Clydesdale, Lachlan; Christopoulos, Arthur; Sexton, Patrick M.; Wootten, Denise

doi:10.1016/j.bcp.2018.09.003

Cited by 50 publications

(52 citation statements)

References 36 publications

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“…Ligand-specific signalling pathway preference ("signal bias") has emerged as a factor controlling downstream GLP-1R actions such as potentiation of insulin secretion (6), and is of ongoing interest in the therapeutic targeting of other membrane receptors as it provides a potential means to accentuate desirable effects and minimise side effects (7). Moreover, endocytosis and post-endocytic trafficking influence the availability of GLP-1Rs at the cell surface and fine-tune the spatiotemporal origin of signalling responses (8)(9)(10).…”

Section: Introductionmentioning

confidence: 99%

The influence of peptide context on signalling and trafficking of glucagon-like peptide-1 receptor biased agonists

Fang

Chen

Pickford

et al. 2020

Preprint

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Signal bias and membrane trafficking have recently emerged as important considerations in the therapeutic targeting of the glucagon-like peptide-1 receptor (GLP-1R) in type 2 diabetes and obesity. In the present study, we have evaluated a peptide series with varying sequence homology between native GLP-1 and exendin-4, the archetypal ligands on which approved GLP-1R agonists are based. We find notable differences in agonist-mediated signalling, endocytosis and recycling, dependent both on the introduction of a His → Phe switch at position 1 and the specific mid-peptide helical regions and C-termini of the two agonists. These observations were linked to insulin secretion in a beta cell model and provide insights into how ligand factors influence GLP-1R function at the cellular level.

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Section: Introductionmentioning

confidence: 99%

The influence of peptide context on signalling and trafficking of glucagon-like peptide-1 receptor biased agonists

Fang

Chen

Pickford

et al. 2020

Preprint

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“…The predominant intracellular signalling intermediate that couples GLP-1R activation to its downstream effects is cyclic adenosine monophosphate (cAMP) (5,6). However, an updated view of GLP-1R pharmacology highlights the roles of membrane trafficking (7,8) and additional effector proteins such as the β-arrestins (9,10) in the control of amplitude, duration and subcellular localisation of signalling events to regulate insulin secretion, particularly in the pharmacological setting. Although all clinically approved GLP-1RAs show broadly similar signalling and trafficking characteristics to the endogenous ligand, GLP-1(7-36)NH2, these can be dramatically altered via sequence modifications close to the ligand N-terminus, as demonstrated recently using analogues of the GLP-1 homologue peptide exendin-4 (11,12).…”

Section: Introductionmentioning

confidence: 99%

Disconnect between signalling potency and in vivo efficacy of pharmacokinetically optimised biased glucagon-like peptide-1 receptor agonists

Lucey

Pickford

Minnion

et al. 2019

Preprint

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AbstractObjectiveTo determine how pharmacokinetically advantageous acylation impacts on glucagon-like peptide-1 receptor (GLP-1R) signal bias, trafficking, anti-hyperglycaemic efficacy and appetite suppression.MethodsIn vitro signalling responses were measured using biochemical and biosensor assays. GLP-1 receptor trafficking was determined by confocal microscopy and diffusion-enhanced resonance energy transfer. Pharmacokinetics, glucoregulatory effects and appetite suppression were measured in acute, sub-chronic and chronic settings in mice.ResultsA C-terminally acylated ligand, exendin-phe1-C16, was identified with undetectable β-arrestin recruitment and GLP-1R internalisation. Depending on the cellular system used, this molecule was up to 1000-fold less potent than the comparator exendin-asp-3-C16 for cyclic AMP signalling, yet was considerably more effective in vivo, particularly for glucose regulation.ConclusionsC-terminal acylation of biased GLP-1R agonists increases their degree of signal bias in favour of cAMP production and improves their therapeutic potential.

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“…With the current sharp rise in the worldwide incidence of both type 2 diabetes and obesity (54), there is a pressing need to develop more effective drugs with fewer associated side effects. The discovery of biased signaling, by which a GPCR can transduce specific signaling profiles by favoring one pathway or behavior over another (55), opens up new avenues to achieve deeper control of GLP-1R action and cellular responses, as previously shown by us (19), and others (56).…”

Section: Discussionmentioning

confidence: 94%

Agonist binding affinity determines palmitoylation of the glucagon-like peptide-1 receptor and its functional interaction with plasma membrane nanodomains in pancreatic beta cells

Buenaventura

Laughlin

Bitsi

et al. 2018

Preprint

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30The glucagon-like peptide-1 receptor (GLP-1R), a key pharmacological target in type 31 2 diabetes and obesity, is known to undergo palmitoylation by covalent ligation of an 32 acyl chain to cysteine 438 in its carboxyl-terminal tail. Work with other GPCRs 33 indicates that palmitoylation can be dynamically regulated to allow receptors to 34 partition into plasma membrane nanodomains that act as signaling hotspots. Here, 35we demonstrate that the palmitoylated state of the GLP-1R is increased by agonist 36 binding, leading to its segregation and clustering into plasma membrane signaling 37 nanodomains before undergoing internalization in a clathrin-dependent manner. Both 38 GLP-1R signaling and trafficking are modulated by strategies targeting nanodomain 39 segregation and cluster formation, including depletion of cholesterol or expression of 40 a palmitoylation-defective GLP-1R mutant. Differences in receptor binding affinity 41 exhibited by biased GLP-1R agonists, and modulation of binding kinetics with the 42 positive allosteric modulator BETP, influence GLP-1R palmitoylation, clustering, 43 nanodomain signaling, and internalization. Downstream effects on insulin secretion 44 from pancreatic beta cells indicate that these processes are relevant to GLP-1R 45 physiological actions and might be therapeutically targetable. 46 47 48 49 50 51 52 53 54 55 56 Introduction 57 58 G protein-coupled receptors (GPCRs), the largest membrane receptor family in 59 eukaryotes (1), are integral membrane proteins and, as such, both their physical 60 organization and their signaling properties are modulated by the lipid composition of 61 the surrounding membrane (2, 3). The localization of GPCRs to dynamic membrane 62nanodomains has been widely reported (4-6). These nanodomains, or membrane 63 rafts, which cannot be directly observed in living cells with current methods (7), are 64 often described as highly organized detergent-resistant, liquid-ordered, 65 glycosphingolipid-and cholesterol-rich platforms where receptor-signaling complexes 66 become compartmentalized, facilitating efficient coupling with G proteins (5, 8, 9). 67Additionally, most GPCRs are modified post-translationally with one or more palmitic 68 acid chains linked covalently, but reversibly, via a thioester bond to cysteines within 69 the intracellular domain of the receptor, in a process known as palmitoylation (2, 10). 70The insertion of acyl chain(s) is regulated by families of acyltransferases (DHHCs) 71 and palmitoyl protein thioesterases (11, 12), and can be either constitutive or 72 modulated by agonist binding (13, 14). GPCR palmitoylation at or near the end of its 73 carboxyl-terminal (C-terminal) tail creates a new membrane anchor and a further 74 intracellular loop (15) that modifies the GPCR structure and its interactions with 75 specific intracellular partners, favoring receptor partitioning into plasma membrane 76 nanodomains (2, 16, 17). 78We have previously described how signaling responses of the glucagon-like peptide-79 1 receptor (GLP-1R), a ...

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Glucagon-like peptide-1 receptor internalisation controls spatiotemporal signalling mediated by biased agonists

Cited by 50 publications

References 36 publications

The influence of peptide context on signalling and trafficking of glucagon-like peptide-1 receptor biased agonists

The influence of peptide context on signalling and trafficking of glucagon-like peptide-1 receptor biased agonists

Disconnect between signalling potency and in vivo efficacy of pharmacokinetically optimised biased glucagon-like peptide-1 receptor agonists

Agonist binding affinity determines palmitoylation of the glucagon-like peptide-1 receptor and its functional interaction with plasma membrane nanodomains in pancreatic beta cells

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