+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-14223 | |||||||||
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Title | Structure of the V2 receptor Cter-arrestin2-ScFv30 complex | |||||||||
Map data | ||||||||||
Sample |
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Function / homology | Function and homology information angiotensin receptor binding / Activation of SMO / negative regulation of interleukin-8 production / arrestin family protein binding / G protein-coupled receptor internalization / enzyme inhibitor activity / Lysosome Vesicle Biogenesis / positive regulation of Rho protein signal transduction / Golgi Associated Vesicle Biogenesis / negative regulation of NF-kappaB transcription factor activity ...angiotensin receptor binding / Activation of SMO / negative regulation of interleukin-8 production / arrestin family protein binding / G protein-coupled receptor internalization / enzyme inhibitor activity / Lysosome Vesicle Biogenesis / positive regulation of Rho protein signal transduction / Golgi Associated Vesicle Biogenesis / negative regulation of NF-kappaB transcription factor activity / stress fiber assembly / negative regulation of Notch signaling pathway / pseudopodium / negative regulation of interleukin-6 production / positive regulation of receptor internalization / clathrin-coated pit / negative regulation of protein ubiquitination / insulin-like growth factor receptor binding / visual perception / Activated NOTCH1 Transmits Signal to the Nucleus / GTPase activator activity / G protein-coupled receptor binding / Signaling by high-kinase activity BRAF mutants / MAP2K and MAPK activation / cytoplasmic vesicle membrane / Signaling by RAF1 mutants / Signaling by moderate kinase activity BRAF mutants / Paradoxical activation of RAF signaling by kinase inactive BRAF / Signaling downstream of RAS mutants / Signaling by BRAF and RAF1 fusions / Thrombin signalling through proteinase activated receptors (PARs) / protein transport / Cargo recognition for clathrin-mediated endocytosis / Clathrin-mediated endocytosis / ubiquitin-dependent protein catabolic process / cytoplasmic vesicle / G alpha (s) signalling events / proteasome-mediated ubiquitin-dependent protein catabolic process / transcription coactivator activity / positive regulation of ERK1 and ERK2 cascade / protein ubiquitination / nuclear body / Ub-specific processing proteases / positive regulation of protein phosphorylation / lysosomal membrane / Golgi membrane / ubiquitin protein ligase binding / chromatin / regulation of transcription by RNA polymerase II / signal transduction / positive regulation of transcription by RNA polymerase II / nucleoplasm / nucleus / plasma membrane / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | Homo sapiens (human) / synthetic construct (others) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.23 Å | |||||||||
Authors | Bous J / Fouillen A / Trapani S / Granier S / Mouillac B / Bron P | |||||||||
Funding support | France, 2 items
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Citation | Journal: Sci Adv / Year: 2022 Title: Structure of the vasopressin hormone-V2 receptor-β-arrestin1 ternary complex. Authors: Julien Bous / Aurélien Fouillen / Hélène Orcel / Stefano Trapani / Xiaojing Cong / Simon Fontanel / Julie Saint-Paul / Joséphine Lai-Kee-Him / Serge Urbach / Nathalie Sibille / Rémy ...Authors: Julien Bous / Aurélien Fouillen / Hélène Orcel / Stefano Trapani / Xiaojing Cong / Simon Fontanel / Julie Saint-Paul / Joséphine Lai-Kee-Him / Serge Urbach / Nathalie Sibille / Rémy Sounier / Sébastien Granier / Bernard Mouillac / Patrick Bron / Abstract: Arrestins interact with G protein-coupled receptors (GPCRs) to stop G protein activation and to initiate key signaling pathways. Recent structural studies shed light on the molecular mechanisms ...Arrestins interact with G protein-coupled receptors (GPCRs) to stop G protein activation and to initiate key signaling pathways. Recent structural studies shed light on the molecular mechanisms involved in GPCR-arrestin coupling, but whether this process is conserved among GPCRs is poorly understood. Here, we report the cryo-electron microscopy active structure of the wild-type arginine-vasopressin V2 receptor (V2R) in complex with β-arrestin1. It reveals an atypical position of β-arrestin1 compared to previously described GPCR-arrestin assemblies, associated with an original V2R/β-arrestin1 interface involving all receptor intracellular loops. Phosphorylated sites of the V2R carboxyl terminus are clearly identified and interact extensively with the β-arrestin1 N-lobe, in agreement with structural data obtained with chimeric or synthetic systems. Overall, these findings highlight a notable structural variability among GPCR-arrestin signaling complexes. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_14223.map.gz | 9.8 MB | EMDB map data format | |
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Header (meta data) | emd-14223-v30.xml emd-14223.xml | 21.1 KB 21.1 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_14223_fsc.xml | 4.6 KB | Display | FSC data file |
Images | emd_14223.png | 89.2 KB | ||
Masks | emd_14223_msk_1.map | 10.5 MB | Mask map | |
Others | emd_14223_additional_1.map.gz emd_14223_half_map_1.map.gz emd_14223_half_map_2.map.gz | 5 MB 9.7 MB 9.7 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-14223 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-14223 | HTTPS FTP |
-Related structure data
Related structure data | 7r0jMC 7r0cC C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_14223.map.gz / Format: CCP4 / Size: 10.5 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.28 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
File | emd_14223_msk_1.map | ||||||||||||
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Density Histograms |
-Additional map: #1
File | emd_14223_additional_1.map | ||||||||||||
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Density Histograms |
-Half map: #2
File | emd_14223_half_map_1.map | ||||||||||||
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Density Histograms |
-Half map: #1
File | emd_14223_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
-Entire : Ternary complex of the Cterminal portion of the V2 receptor with ...
Entire | Name: Ternary complex of the Cterminal portion of the V2 receptor with arrestin2 and ScfV30 |
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Components |
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-Supramolecule #1: Ternary complex of the Cterminal portion of the V2 receptor with ...
Supramolecule | Name: Ternary complex of the Cterminal portion of the V2 receptor with arrestin2 and ScfV30 type: complex / Chimera: Yes / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: Homo sapiens (human) |
-Macromolecule #1: V2R Cter
Macromolecule | Name: V2R Cter / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 1.780248 KDa |
Recombinant expression | Organism: Insect expression vector pBlueBacmsGCA1 (others) |
Sequence | String: E(SEP)C(TPO)(TPO)A(SEP)(SEP)(SEP)L AKD |
-Macromolecule #2: Arrestin2
Macromolecule | Name: Arrestin2 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 47.164609 KDa |
Recombinant expression | Organism: Escherichia coli (E. coli) |
Sequence | String: MGASWSHPQF EKGGGSGGGS GGSSAWSHPQ FEKLEVLFQG PASGDKGTRV FKKASPNGKL TVYLGKRDFV DHIDLVDPVD GVVLVDPEY LKERRVYVTL TCAFRYGRED LDVLGLTFRK DLFVANVQSF PPAPEDKKPL TRLQERLIKK LGEHAYPFTF E IPPNLPCS ...String: MGASWSHPQF EKGGGSGGGS GGSSAWSHPQ FEKLEVLFQG PASGDKGTRV FKKASPNGKL TVYLGKRDFV DHIDLVDPVD GVVLVDPEY LKERRVYVTL TCAFRYGRED LDVLGLTFRK DLFVANVQSF PPAPEDKKPL TRLQERLIKK LGEHAYPFTF E IPPNLPCS VTLQPGPEDT GKACGVDYEV KAFCAENLEE KIHKRNSVRL VIRKVQYAPE RPGPQPTAET TRQFLMSDKP LH LEASLDK EIYYHGEPIS VNVHVTNNTN KTVKKIKISV RQYADICLFN TAQYKCPVAM EEADDTVAPS STFCKVYTLT PFL ANNREK RGLALDGKLK HEDTNLASST LLREGANREI LGIIVSYKVK VKLVVSRGGL LGDLASSDVA VELPFTLMHP KPKE EPPHR EVPENETPVD TNLIELDTN |
-Macromolecule #3: ScFv30
Macromolecule | Name: ScFv30 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: synthetic construct (others) |
Molecular weight | Theoretical: 30.070027 KDa |
Recombinant expression | Organism: Insect expression vector pBlueBacmsGCA1 (others) |
Sequence | String: AMGDIQMTQS PSSLSASVGD RVTITCRASQ SVSSAVAWYQ QKPGKAPKLL IYSASSLYSG VPSRFSGSRS GTDFTLTISS LQPEDFATY YCQQYKYVPV TFGCGTKVEI KGTTAASGSS GGSSSGAEVQ LVESGGGLVQ PGGSLRLSCA ASGFNVYSSS I HWVRQAPG ...String: AMGDIQMTQS PSSLSASVGD RVTITCRASQ SVSSAVAWYQ QKPGKAPKLL IYSASSLYSG VPSRFSGSRS GTDFTLTISS LQPEDFATY YCQQYKYVPV TFGCGTKVEI KGTTAASGSS GGSSSGAEVQ LVESGGGLVQ PGGSLRLSCA ASGFNVYSSS I HWVRQAPG KCLEWVASIS SYYGYTYYAD SVKGRFTISA DTSKNTAYLQ MNSLRAEDTA VYYCARSRQF WYSGLDYWGQ GT LVTVSSA AADDDDKAGW SHPQFEKGGG SGGGSGGGSW SHPQFEK |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 3 mg/mL |
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Buffer | pH: 7.5 |
Grid | Model: UltrAuFoil R1.2/1.3 / Material: GOLD / Mesh: 300 / Support film - Material: GOLD / Support film - topology: CONTINUOUS / Pretreatment - Type: GLOW DISCHARGE |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: LEICA EM GP |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 50.0 µm / Calibrated magnification: 130000 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / Nominal defocus max: 2.0 µm / Nominal defocus min: 1.0 µm |
Sample stage | Cooling holder cryogen: NITROGEN |
Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Number real images: 14080 / Average exposure time: 1.0 sec. / Average electron dose: 52.0 e/Å2 |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |