Molecular Basis for Hormone Recognition and Activation of Corticotropin-Releasing Factor Receptors

Ma, Shanshan; Shen, Qingya; Zhao, Longfeng; Mao, Chunyou; Zhou, Xin; Shen, Dan; Waal, Parker W. de; Bi, Peng; Li, Chuntao; Jiang, Yi; Wang, Ming Wei; Sexton, Patrick M.; Wootten, Denise; Melcher, Karsten; Zhang, Yan; Xu, Hao

doi:10.1016/j.molcel.2020.01.013

Cited by 77 publications

(102 citation statements)

References 53 publications

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“…For GCGRapo, the motions of the ECD accounted for by the Fig. S3), and the importance of ECD dynamics has been stressed in a number of studies (2,13,57,58). Our simulations further reveal that different conformations of ECD have different dynamic behaviour which may have functional relevance, e.g.…”

Section: ) Ofsupporting

confidence: 51%

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The Glycosphingolipid GM3 Modulates Conformational Dynamics of the Glucagon Receptor

Ansell

Song

Sansom

2020

Preprint

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The extracellular domain (ECD) of Class B1 G-protein coupled receptors (GPCRs) plays a central role in signal transduction and is uniquely positioned to sense both the extracellular and membrane environments. Whilst recent studies suggest a role for membrane lipids in the modulation of Class A and Class F GPCR signalling properties, little is known about the effect of lipids on Class B1 receptors. In this study, we employed multiscale molecular dynamics (MD) simulations to access the dynamics of the glucagon receptor (GCGR) ECD in the presence of native-like membrane bilayers.Simulations showed that the ECD could move about a hinge region formed by residues Q122-E126 to adopt both closed and open conformations relative to the TMD. ECD movements were modulated by binding of the glycosphingolipid GM3. These large-scale fluctuations in ECD conformation that may affect the ligand binding and receptor activation properties. We also identify a unique PIP2 interaction profile near ICL2/TM3 at the G-protein coupling interface, suggesting a mechanism of engaging G-proteins which may have a distinct dependence on PIP2 compared to Class A GPCRs.Given the structural conservation of Class B1 GPCRs, the modulatory effects of GM3 and PIP2 on GCGR may be conserved across these receptors, offering new insights into potential therapeutic targeting. Statement of SignificanceThe role of lipids in regulation of Class B GPCRs remains elusive, despite recent structural advances. In this study, multi-scale molecular dynamics simulations are used to evaluate lipid interactions with the glucagon receptor, a Class B1 GPCR. We find that the glycosphingolipid GM3 binds to the glucagon receptor extracellular domain (ECD), modulating the dynamics of the ECD and promoting movement away from the transmembrane domain . We also identify a unique PIP2 interaction fingerprint in a region known to be important for bridging G-protein coupling in Class A GPCRs. Thus, this study provides molecular insight into the behaviour of the glucagon receptor in a complex lipid bilayer environment which may aid understanding of glucagon receptor signalling properties.GCGR_main text v16 figs MS biorxiv.docx

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Section: ) Ofsupporting

confidence: 51%

“…The ECD of GCGRapo has a distinct conformation compared to that of GCGRpept which represents the canonical peptide-bound conformation as seen in several Class B1 GPCRs (Supporting Material Fig. S3) (51)(52)(53)(54)(55)(56)(57)(58). The stalk in GCGRapo also forms a β-sheet with ECL1.…”

Section: Open and Closed Conformations Of The Ecdmentioning

confidence: 99%

The Glycosphingolipid GM3 Modulates Conformational Dynamics of the Glucagon Receptor

Ansell

Song

Sansom

2020

Preprint

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“…These receptors predominantly couple to the stimulatory protein Gs to promote cAMP production, however they can activate a range of signalling pathways and are highly susceptible to biased agonism (Wootten et al, 2017). In the past three years numerous active state structures of class B1 GPCRs coupled to the stimulatory protein Gs have been determined using single particle cryo-electron microscopy (cryo-EM) (Liang et al, 2020a;Liang et al, 2020b;Liang et al, 2018a;Liang et al, 2018b;Liang et al, 2017;Ma et al, 2020;Qiao et al, 2020;Zhang et al, 2017;Zhao et al, 2019). The resulting structures revealed common peptide binding pockets and a conserved Gs binding site within the receptor transmembrane (TM) helix bundle.…”

Section: Introductionmentioning

confidence: 99%

Differential GLP-1R binding and activation by peptide and non-peptide agonists

Zhang

Belousoff

Zhao

et al. 2020

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SUMMARYPeptide drugs targeting class B1 GPCRs can treat multiple diseases, however there remains substantial interest in the development of orally delivered non-peptide drugs. Here we reveal unexpected overlap between signalling and regulation of the glucagon-like peptide-1 (GLP-1) receptor by the non-peptide agonist, PF 06882961, and GLP-1 that was not observed for another compound, OWL-833. Both compounds are currently in clinical trials for treatment of type 2 diabetes. High resolution cryo-EM structures reveal the binding sites for PF-06882961 and GLP-1 substantially overlap, whereas OWL-833 adopts a unique binding mode with a more open receptor conformation at the extracellular face. Structural differences involving extensive water-mediated hydrogen bond networks could be correlated to functional data to understand how PF 06882961, but not OWL-833, can closely mimic the pharmacological properties of GLP-1. These findings will facilitate rational structure-based discovery of non-peptide agonists targeting class B GPCRs.

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“…Thus, our structures provide accurate models of the full-length GABA B receptor in the inactive and active states, shed lights on intersubunit interactions between GB1 and GB2, ligand recognition and proposed models of G i1 engagement to the receptor. We also observed multiply ordered cholesterols not only surrounding the periphery of both TM domains like many other GPCRs 37,38 , but also between two subunits primarily in the inactive state (Extended Data Fig. 6a).…”

Section: Resultsmentioning

confidence: 67%

Cryo-EM structures of inactive and G_i-coupled GABA_B heterodimer

Mao

Shen

et al. 2020

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22Metabotropic GABA B G protein-coupled receptor functions as a mandatory heterodimer of 23 GB1 and GB2 subunits and mediates inhibitory neurotransmission in the central nervous system. 24Each subunit is composed of the extracellular Venus flytrap (VFT) domain and transmembrane 25 (TM) domain. Here we present cryo-EM structures of human full-length heterodimeric GABA B 26 receptor in the antagonist-bound inactive state and in the active state complexed with agonist 27 and positive allosteric modulator in the presence of G i1 protein at a resolution range of 2.8-3.0 28 Å. Cryo-EM analysis of the activated-GABA B -G i1 complex revealed that G i1 couples to the 29 activated receptor primarily in three major conformations, one via GB1 TM and two via GB2 30 TM, respectively. Our structures reveal that agonist binding stabilizes the closure of GB1 VFT, 31 which in turn triggers a rearrangement of TM interfaces between two subunits from TM3-32 TM5/TM3-TM5 in the inactive state to TM6/TM6 in the active state and finally induces the 33 opening of intracellular loop 3 and synergistically shifting of TM3, 4 and 5 helices in GB2 TM 34 domain to accommodate the α5-helix of G i1 . These results provide a structural framework for 35 understanding class C GPCR activation and a rational template for allosteric modulator design 36 targeting dimeric interface of GABA B receptor. 37 38 GABA B receptor belongs to the class C GPCR composed by metabotropic glutamate 54 receptors (mGlus), calcium-sensing receptor (CaSR) and taste 1 receptors 16 . Class C GPCR 55 form obligatory dimers, among them, mGlus and CaSR form the homodimer 17,18 , whereas 56 GABA B receptor is an obligatory heterodimer of two subunits GABA B1 (GB1) and GABA B2 57 (GB2) 19,20 . Each subunit is composed of a large extracellular 'Venus Flytrap' (VFT) domain, a 58 transmembrane (TM) domain, and a cytoplasmic tail 21 (Extended Data Fig. 1a, b). GB1 59 possesses an endoplasmic reticulum (ER) retention motif in its cytoplasmic tail 22 . The 60interaction between GB1 and GB2 facilitates its cell surface expression through coiled-coil 61 interactions in its cytoplasmic tail 23 . While GB1 is responsible for ligand recognition through 62 its VFT 24 , GB2 couples G i/o proteins through its TM 25,26 . However, a line of evidence show that 63 in the absence of GB2, GB1 asa (a GB1 mutant with deletion of ER retention signal) at the cell 64 surface or ER-localized GB1 also induces downstream signaling through G i/o proteins 27,28 . 65The crystal structures of isolated heterodimeric VFT of GABA B receptor in presence of the 66 antagonist or agonist revealed the open or closed conformation of GB1 VFT 29 . Structures of 67 isolated VFT and TM domains from other class C GPCR have been reported [30][31][32] , in addition to 68 more recently breakthrough that cryo-EM structures of full-length mGlu5 in apo state and in 69 the presence of agonist have been determined both at overall 4 Å resolution, providing the first 70 insights into the architecture of mGlu5 and structural framew...

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Molecular Basis for Hormone Recognition and Activation of Corticotropin-Releasing Factor Receptors

Cited by 77 publications

References 53 publications

The Glycosphingolipid GM3 Modulates Conformational Dynamics of the Glucagon Receptor

The Glycosphingolipid GM3 Modulates Conformational Dynamics of the Glucagon Receptor

Differential GLP-1R binding and activation by peptide and non-peptide agonists

Cryo-EM structures of inactive and G_i-coupled GABA_B heterodimer

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Molecular Basis for Hormone Recognition and Activation of Corticotropin-Releasing Factor Receptors

Cited by 77 publications

References 53 publications

The Glycosphingolipid GM3 Modulates Conformational Dynamics of the Glucagon Receptor

The Glycosphingolipid GM3 Modulates Conformational Dynamics of the Glucagon Receptor

Differential GLP-1R binding and activation by peptide and non-peptide agonists

Cryo-EM structures of inactive and Gi-coupled GABAB heterodimer

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Cryo-EM structures of inactive and G_i-coupled GABA_B heterodimer