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Yorodumi- EMDB-17345: Cryo-EM structure of Rhodopsin-Gi bound with antibody fragments s... -
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Basic information
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| Title | Cryo-EM structure of Rhodopsin-Gi bound with antibody fragments scFv16 and Fab79, conformation 2 | |||||||||||||||
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Keywords | GPCR / Rhodopsin / G protein / Fab / SIGNALING PROTEIN | |||||||||||||||
| Function / homology | Function and homology informationOpsins / VxPx cargo-targeting to cilium / rod bipolar cell differentiation / sperm head plasma membrane / opsin binding / The canonical retinoid cycle in rods (twilight vision) / absorption of visible light / G protein-coupled opsin signaling pathway / Olfactory Signaling Pathway / Sensory perception of sweet, bitter, and umami (glutamate) taste ...Opsins / VxPx cargo-targeting to cilium / rod bipolar cell differentiation / sperm head plasma membrane / opsin binding / The canonical retinoid cycle in rods (twilight vision) / absorption of visible light / G protein-coupled opsin signaling pathway / Olfactory Signaling Pathway / Sensory perception of sweet, bitter, and umami (glutamate) taste / 11-cis retinal binding / podosome assembly / Synthesis, secretion, and inactivation of Glucagon-like Peptide-1 (GLP-1) / G protein-coupled photoreceptor activity / photoreceptor inner segment membrane / cellular response to light stimulus / rod photoreceptor outer segment / eye photoreceptor cell development / G protein-coupled receptor complex / Inactivation, recovery and regulation of the phototransduction cascade / thermotaxis / Activation of the phototransduction cascade / outer membrane / detection of temperature stimulus involved in thermoception / response to light intensity / photoreceptor cell maintenance / arrestin family protein binding / Activation of G protein gated Potassium channels / G-protein activation / G beta:gamma signalling through PI3Kgamma / Prostacyclin signalling through prostacyclin receptor / G beta:gamma signalling through PLC beta / ADP signalling through P2Y purinoceptor 1 / Thromboxane signalling through TP receptor / Presynaptic function of Kainate receptors / G beta:gamma signalling through CDC42 / Inhibition of voltage gated Ca2+ channels via Gbeta/gamma subunits / G alpha (12/13) signalling events / Glucagon-type ligand receptors / G beta:gamma signalling through BTK / ADP signalling through P2Y purinoceptor 12 / Adrenaline,noradrenaline inhibits insulin secretion / Cooperation of PDCL (PhLP1) and TRiC/CCT in G-protein beta folding / Ca2+ pathway / G alpha (z) signalling events / Thrombin signalling through proteinase activated receptors (PARs) / Extra-nuclear estrogen signaling / G alpha (s) signalling events / G alpha (q) signalling events / photoreceptor outer segment membrane / G alpha (i) signalling events / Glucagon-like Peptide-1 (GLP1) regulates insulin secretion / High laminar flow shear stress activates signaling by PIEZO1 and PECAM1:CDH5:KDR in endothelial cells / Vasopressin regulates renal water homeostasis via Aquaporins / phototransduction, visible light / phototransduction / response to light stimulus / photoreceptor outer segment / G-protein alpha-subunit binding / adenylate cyclase inhibitor activity / positive regulation of protein localization to cell cortex / T cell migration / positive regulation of relaxation of smooth muscle / Adenylate cyclase inhibitory pathway / D2 dopamine receptor binding / visual perception / adenylate cyclase-inhibiting serotonin receptor signaling pathway / G protein-coupled serotonin receptor binding / guanyl-nucleotide exchange factor activity / cellular response to forskolin / regulation of mitotic spindle organization / chemokine-mediated signaling pathway / neuropeptide signaling pathway / Regulation of insulin secretion / response to prostaglandin E / positive regulation of cholesterol biosynthetic process / negative regulation of insulin secretion / G protein-coupled receptor binding / response to peptide hormone / microtubule cytoskeleton organization / adenylate cyclase-modulating G protein-coupled receptor signaling pathway / G-protein beta/gamma-subunit complex binding / centriolar satellite / adenylate cyclase-inhibiting G protein-coupled receptor signaling pathway / cell-cell junction / ADP signalling through P2Y purinoceptor 12 / photoreceptor disc membrane / Adrenaline,noradrenaline inhibits insulin secretion / GDP binding / intracellular protein localization / G alpha (z) signalling events / ADORA2B mediated anti-inflammatory cytokines production / cellular response to catecholamine stimulus / adenylate cyclase-activating dopamine receptor signaling pathway / GPER1 signaling / cellular response to prostaglandin E stimulus / G-protein beta-subunit binding / heterotrimeric G-protein complex / sensory perception of taste / sperm principal piece Similarity search - Function | |||||||||||||||
| Biological species | ![]() Homo sapiens (human) / ![]() | |||||||||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 5.9 Å | |||||||||||||||
Authors | Pamula F / Tejero O / Muehle J / Thoma R / Schertler GFX / Marino J / Tsai C-J | |||||||||||||||
| Funding support | Switzerland, European Union, 4 items
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Citation | Journal: Biophys J / Year: 2025Title: Tool antibody fragments reveal multiple conformations of the rhodopsin-Gi signaling complex. Authors: Filip Pamula / Oliver Tejero / Jonas Mühle / Ralf Thoma / Gebhard F X Schertler / Jacopo Marino / Ching-Ju Tsai / ![]() Abstract: Antibody Fab fragments are widely used protein binders that assist in structural studies of G-protein-coupled receptor (GPCR) signaling complexes. Expanding the repertoire of such binders to target ...Antibody Fab fragments are widely used protein binders that assist in structural studies of G-protein-coupled receptor (GPCR) signaling complexes. Expanding the repertoire of such binders to target distinct components of the signaling complex offers opportunities to probe conformational regulation and dynamics. Here, we report the biochemical and cryo-EM characterization of two Fab fragments, Fab79 and Fab13, raised against the rhodopsin-Gαiβγ complex. Fab79 binds to the flexible α-helical domain (AHD) of the Gαi subunit and prevents complex dissociation in the presence of the nonhydrolyzable GTP analog, GTPγS, likely by hindering AHD closure, a step necessary for complex dissociation. In contrast, Fab13 binds rigidly to Gβ without directly contacting Gα or the receptor. These findings show that Fab79 and Fab13 reveal functionally relevant conformational states of G-protein activation and serve as practical tools to stabilize or modulate GPCR signaling complexes. | |||||||||||||||
| History |
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Structure visualization
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Downloads & links
-EMDB archive
| Map data | emd_17345.map.gz | 18.1 MB | EMDB map data format | |
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| Header (meta data) | emd-17345-v30.xml emd-17345.xml | 33.4 KB 33.4 KB | Display Display | EMDB header |
| Images | emd_17345.png | 68.3 KB | ||
| Filedesc metadata | emd-17345.cif.gz | 8.4 KB | ||
| Others | emd_17345_half_map_1.map.gz emd_17345_half_map_2.map.gz | 165.8 MB 165.8 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-17345 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-17345 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 8p15MC ![]() 8p12C ![]() 8p13C C: citing same article ( M: atomic model generated by this map |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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Map
| File | Download / File: emd_17345.map.gz / Format: CCP4 / Size: 209.3 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: 0.854 Å | ||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
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-Supplemental data
-Half map: #1
| File | emd_17345_half_map_1.map | ||||||||||||
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| Density Histograms |
-Half map: #2
| File | emd_17345_half_map_2.map | ||||||||||||
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| Density Histograms |
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Sample components
+Entire : Rhodopsin-Gi-scFv16-Fab79 complex
+Supramolecule #1: Rhodopsin-Gi-scFv16-Fab79 complex
+Supramolecule #2: Rhodopsin
+Supramolecule #3: Guanine nucleotide-binding protein G(i) subunit alpha-1
+Supramolecule #4: Guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-1 ...
+Supramolecule #5: single-chain Fv scFv16
+Supramolecule #6: Immunoglobin G heavy chain FAB fragment and Immunoglobin G light chain
+Macromolecule #1: Rhodopsin
+Macromolecule #2: Guanine nucleotide-binding protein G(i) subunit alpha-1
+Macromolecule #3: Guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-1
+Macromolecule #4: Guanine nucleotide-binding protein G(T) subunit gamma-T1
+Macromolecule #5: single-chain Fv scFv16
+Macromolecule #6: Immunoglobin G heavy chain FAB fragment
+Macromolecule #7: Immunoglobin G light chain
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Concentration | 0.7 mg/mL | |||||||||
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| Buffer | pH: 7.5 Component:
Details: 20 mM HEPES (pH 7.5), 100 mM NaCl | |||||||||
| Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 200 / Pretreatment - Type: GLOW DISCHARGE | |||||||||
| Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV | |||||||||
| Details | Monodisperse |
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Electron microscopy
| Microscope | FEI TITAN KRIOS |
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| Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average electron dose: 60.0 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.4 µm / Nominal defocus min: 0.8 µm / Nominal magnification: 165000 |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
| Initial model |
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| Refinement | Space: REAL / Protocol: AB INITIO MODEL / Overall B value: 295 / Target criteria: cross-correlation coefficient | ||||||||||||||||||||||||||||||
| Output model | ![]() PDB-8p15: |
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About Yorodumi



Keywords
Homo sapiens (human)
Authors
Switzerland, European Union, 4 items
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Y (Row.)
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Trichoplusia ni (cabbage looper)
FIELD EMISSION GUN


