Cryo-EM structure of an activated VIP1 receptor-G protein complex revealed by a NanoBiT tethering strategy

Duan, Jia; Shen, Dan‐Dan; Zhou, Xuemei; Bi, Peng; Liu, Qiu-feng; Tan, Yangxia; Zhuang, Yahui; Zhang, Hui-bing; Xu, Peiyu; Huang, Sijie; Ma, Shanshan; He, Xinheng; Melcher, Karsten; Zhang, Yan; Xu, Hao; Jiang, Yi

doi:10.1038/s41467-020-17933-8

Cited by 177 publications

(214 citation statements)

References 51 publications

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“…We used NanoBiT strategy 22 – 24 to stabilize the complex, in which GLP-2R was directly attached to LgBiT and rat Gβ was linked to HiBiT with a 15-amino acid linker in between. Transiently transfected HEK 293T cells showed that the C-terminus-truncated GLP-2R(1–490)-LgBiT had a 6-fold stronger interaction with Gβ-SmBiT than the wild-type (WT) receptor(1–553)-LgBiT (Supplementary information, Fig.…”

Section: Resultsmentioning

confidence: 99%

“…In this study, we employed cryo-EM to determine the high-resolution structure of the human GLP-2R in complex with G s protein. Based on our prior experience, 22 NanoBiT strategy 23 , 24 was used in this study to stabilize the GLP-2–GLP-2R–G s complex and strengthen the interactions between GLP-2R and Gβ, resulting in a cryo-EM structure at a global resolution of 3.0 Å (Supplementary information, Table S1 ). Our data provide a rational template to facilitate the design of better GLP-2 analogues for therapeutic use and expand our knowledge on the biology of this receptor family.…”

Section: Introductionmentioning

confidence: 99%

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A unique hormonal recognition feature of the human glucagon-like peptide-2 receptor

et al. 2020

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Glucagon-like peptides (GLP-1 and GLP-2) are two proglucagon-derived intestinal hormones that mediate distinct physiological functions through two related receptors (GLP-1R and GLP-2R) which are important drug targets for metabolic disorders and Crohn’s disease, respectively. Despite great progress in GLP-1R structure determination, our understanding on the differences of peptide binding and signal transduction between these two receptors remains elusive. Here we report the electron microscopy structure of the human GLP-2R in complex with GLP-2 and a Gs heterotrimer. To accommodate GLP-2 rather than GLP-1, GLP-2R fine-tunes the conformations of the extracellular parts of transmembrane helices (TMs) 1, 5, 7 and extracellular loop 1 (ECL1). In contrast to GLP-1, the N-terminal histidine of GLP-2 penetrates into the receptor core with a unique orientation. The middle region of GLP-2 engages with TM1 and TM7 more extensively than with ECL2, and the GLP-2 C-terminus closely attaches to ECL1, which is the most protruded among 9 class B G protein-coupled receptors (GPCRs). Functional studies revealed that the above three segments of GLP-2 are essential for GLP-2 recognition and receptor activation, especially the middle region. These results provide new insights into the molecular basis of ligand specificity in class B GPCRs and may facilitate the development of more specific therapeutics.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A unique hormonal recognition feature of the human glucagon-like peptide-2 receptor

et al. 2020

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“…6d, e ). 9 , 108 The binding of a peptidic ligand causes a conformation rearrangement of the central polar network with simultaneous destabilization of the TM6 helix, thus initiating a sharp kink formation at the conserved motif P 6.47b xxG 6.50b . This central polar network is preserved across the class B receptors solved so far.…”

Section: Gpcr Structurementioning

confidence: 99%

G protein-coupled receptors: structure- and function-based drug discovery

Yang

Zhou

Labroska

et al. 2021

Sig Transduct Target Ther

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As one of the most successful therapeutic target families, G protein-coupled receptors (GPCRs) have experienced a transformation from random ligand screening to knowledge-driven drug design. We are eye-witnessing tremendous progresses made recently in the understanding of their structure–function relationships that facilitated drug development at an unprecedented pace. This article intends to provide a comprehensive overview of this important field to a broader readership that shares some common interests in drug discovery.

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“…G-protein-coupled receptors (GPCRs) are the largest and most diverse group of membrane receptors in eukaryotes and play a role in an incredible array of functions in the human body. 19 …”

Section: Discussionmentioning

confidence: 99%

GPR120 Agonist GW9508 Ameliorated Cellular Senescence Induced by ox-LDL

Liu

Cheng²,

Zeng

et al. 2020

ACS Omega

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Introduction Oxidized low-density lipoprotein (ox-LDL)-induced endothelial senescence is involved in the pathogenesis of atherosclerosis and many cardiovascular diseases. G-protein-coupled receptor 120 (GPR120), a type of orphan G-protein-coupled receptors (GPRs), plays a vital role in mediating anti-inflammatory and insulin-sensitizing effects. The biological function of GPR120 in vascular endothelial cells is largely unknown. Methods The human aortic endothelial cells (HAECs) were treated with ox-LDL (100 μg/mL) in the presence or absence of GW9508 (50 μM) or AH9614 (1 μM) for 24 h. The LDH assay was used to determine cell death. The dihydroethidium (DHE) staining assay was used to measure intracellular levels of reactive oxidative species (ROS), and a senescence β-galactosidase assay kit was used to determine endothelial senescence. Gene and protein expressions were measured using real-time polymerase chain reaction (PCR) and western blot analysis, respectively. Results Ox-LDL treatment decreased the expression of GPR120 by more than half in HAECs. Typically, 100 μg/mL of ox-LDL- induced 35.2% LDH release, which was reduced to 16.9% by 50 μM GW9508, the agonist of GPR120. Importantly, GW9508 relieved cytotoxicity and suppressed the ox-LDL-induced increase in the activity of senescence-associated β-galactosidase (SA-β-Gal) (from 3.3-fold to 1.6-fold of the control group) and the generation of cellular reactive oxidative species (ROS) (from 3.8-fold to 1.6-fold of the control group). Furthermore, we found that GW9508 ameliorated ox-LDL-induced endothelial cell cycle arrest at the G0/G1 phase and the expression of key senescence proteins, including p53 and plasminogen activator inhibitor-1(PAI-1). Mechanistically, we showed that GW9508 promoted ox-LDL-induced transcriptional factor NF-E2-related factor 2 (NRF2) (increase by 47.3%) translocation into the nucleus. The effect of GW9508 is dependent on its receptor GPR120, the blockage of which by its specific antagonist, AH7614, abolished the antisenescence effect of GW9508. Conclusion Collectively, this study revealed the protective effect of GPR120 activation in vascular endothelial cells, implying that GPR120 is a promising therapeutic target for the treatment of cardiovascular diseases.

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Cryo-EM structure of an activated VIP1 receptor-G protein complex revealed by a NanoBiT tethering strategy

Cited by 177 publications

References 51 publications

A unique hormonal recognition feature of the human glucagon-like peptide-2 receptor

A unique hormonal recognition feature of the human glucagon-like peptide-2 receptor

G protein-coupled receptors: structure- and function-based drug discovery

GPR120 Agonist GW9508 Ameliorated Cellular Senescence Induced by ox-LDL

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