Structural insights into probe-dependent positive allosterism of the GLP-1 receptor

Bueno, Ana B.; Sun, Bingfa; Willard, Francis S.; Feng, Dan; Ho, Joseph; Wainscott, David B.; Showalter, Aaron D.; Vieth, Michał; Chen, Qi; Stutsman, Cynthia; Chau, B. Nelson; Ficorilli, James; Agejas, Francisco J; Cumming, Graham R.; Jiménez, Alma; Rojo, Isabel; Kobilka, Tong Sun; Kobilka, Brian K.; Sloop, Kyle W.

doi:10.1038/s41589-020-0589-7

Cited by 72 publications

(88 citation statements)

References 32 publications

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“…In Vitro Pharmacology. cAMP accumulation, β-arrestin recruitment, and receptor binding assays were performed as previously described (28,40).…”

Section: Methodsmentioning

confidence: 99%

“…GLP-1R/LY3502970/G siN18 complex formation and purification. The GLP-1R/LY3502970/ G siN18 complex (referred to as "the complex" in the following text) was formed on membrane as previously described (2,40) with slight modification. The main difference was that we used a modified version of the alpha subunit of the stimulatory G protein, namely, G siN18 , which has the 1 to 18 residues of G ai at its N terminus, replacing the original 1 to 25 residues of G as .…”

Section: Methodsmentioning

confidence: 99%

“…Cryo-EM data acquisition and processing. This procedure was carried out as previously described (40). A sample of 3.5 μL of purified GLP-1R/LY3502970/ G siN18 complex at a concentration of ∼10 mg/mL was applied to glowdischarged holey carbon grids (Quantifoil R1.2/1.3, 200 mesh) and subsequently vitrified using a Vitrobot Mark IV (Thermo Fisher Scientific).…”

Section: Methodsmentioning

confidence: 99%

“…Model Building and Refinement. This procedure was carried out as previously described (40). An initial model was formed by rigid-body fitting of the reported GLP-1R/GLP-1/G s /Nb35 structure (Research Collaboratory for Structural Bioinformatics Protein Data Bank [PDB] ID code 5VAI).…”

Section: Methodsmentioning

confidence: 99%

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Structural basis for GLP-1 receptor activation by LY3502970, an orally active nonpeptide agonist

Kawai¹,

Sun

Yoshino³

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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Glucagon-like peptide-1 receptor (GLP-1R) agonists are efficacious antidiabetic medications that work by enhancing glucose-dependent insulin secretion and improving energy balance. Currently approved GLP-1R agonists are peptide based, and it has proven difficult to obtain small-molecule activators possessing optimal pharmaceutical properties. We report the discovery and mechanism of action of LY3502970 (OWL833), a nonpeptide GLP-1R agonist. LY3502970 is a partial agonist, biased toward G protein activation over β-arrestin recruitment at the GLP-1R. The molecule is highly potent and selective against other class B G protein–coupled receptors (GPCRs) with a pharmacokinetic profile favorable for oral administration. A high-resolution structure of LY3502970 in complex with active-state GLP-1R revealed a unique binding pocket in the upper helical bundle where the compound is bound by the extracellular domain (ECD), extracellular loop 2, and transmembrane helices 1, 2, 3, and 7. This mechanism creates a distinct receptor conformation that may explain the partial agonism and biased signaling of the compound. Further, interaction between LY3502970 and the primate-specific Trp33 of the ECD informs species selective activity for the molecule. In efficacy studies, oral administration of LY3502970 resulted in glucose lowering in humanized GLP-1R transgenic mice and insulinotropic and hypophagic effects in nonhuman primates, demonstrating an effect size in both models comparable to injectable exenatide. Together, this work determined the molecular basis for the activity of an oral agent being developed for the treatment of type 2 diabetes mellitus, offering insights into the activation of class B GPCRs by nonpeptide ligands.

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“…In Vitro Pharmacology. cAMP accumulation, β-arrestin recruitment, and receptor binding assays were performed as previously described (28,40).…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Structural basis for GLP-1 receptor activation by LY3502970, an orally active nonpeptide agonist

Kawai¹,

Sun

Yoshino³

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

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“…PAMs can display various effects (Christopoulos 2014), including increasing agonist potency (from 2 to 100 fold), increasing agonist efficacy, partially activating receptors (ago-PAM effect) or even orienting the receptor towards one of its signaling pathway (Rook et al 2013; Makita et al 2007). It is commonly considered that PAMs act by stabilizing a specific conformation of the receptor (Shaye et al 2020; Bueno et al 2020; Kruse et al 2013; Liu et al 2019; Srivastava et al 2014). However, PAMs may likely act by influencing the equilibrium between preexisting GPCR conformational states.…”

Section: Introductionmentioning

confidence: 99%

Allosteric modulators enhance agonist efficacy by increasing the residence time of a GPCR in the active state

Cao

Quast

Fatemi

et al. 2021

Preprint

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Metabotropic glutamate receptors (mGlu) are G protein-coupled receptors that represent promising targets for brain diseases. Much hope in drug development come from the discovery of positive allosteric modulators that display subtype selectivity, and act by increasing agonist potency, as well as efficacy in most cases. How such compounds can influence agonist efficacy remains unclear. Here, we explore the structural dynamics of the full-length mGlu2 dimers at submillisecond timescales using single molecule FRET on diffusing receptors in optimized detergent micelles. We show that glutamate binding in the Venus flytrap extracellular domains does not stabilize fully the receptors in their active states. The full activation of all receptors can only be observed in the presence of either a positive allosteric modulator or the Gi protein. Our results provide important new insights on the fast kinetics and the action of the allosteric modulators on mGlu activation at the single molecule level.

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Rationalizing the Unprecedented Stereochemistry of an Enzymatic Nitrile Synthesis through a Combined Computational and Experimental Approach

2021

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In this contribution, the unique and unprecedented stereochemical phenomenon of an aldoxime dehydratase‐catalyzed enantioselective dehydration of racemic E‐ and Z‐aldoximes with selective formation of both enantiomeric forms of a chiral nitrile is rationalized by means of molecular modelling, comprising in silico mutations and docking studies. This theoretical investigation gave detailed insight into why with the same enzyme the use of racemic E‐ and Z‐aldoximes leads to opposite forms of the chiral nitrile. The calculated mutants with a larger or smaller cavity in the active site were then prepared and used in biotransformations, showing the theoretically predicted decrease and increase of the enantioselectivities in these nitrile syntheses. This validated model also enabled the rational design of mutants with a smaller cavity, which gave superior enantioselectivities compared to the known wild‐type enzyme, with excellent E‐values of up to E>200 when the mutant OxdRE‐Leu145Phe was utilized.

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Structural insights into probe-dependent positive allosterism of the GLP-1 receptor

Cited by 72 publications

References 32 publications

Structural basis for GLP-1 receptor activation by LY3502970, an orally active nonpeptide agonist

Structural basis for GLP-1 receptor activation by LY3502970, an orally active nonpeptide agonist

Allosteric modulators enhance agonist efficacy by increasing the residence time of a GPCR in the active state

Rationalizing the Unprecedented Stereochemistry of an Enzymatic Nitrile Synthesis through a Combined Computational and Experimental Approach

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