National Institutes of Health/National Cancer Institute (NIH/NCI)
R01 CA201402
United States
National Institutes of Health/National Institute of Neurological Disorders and Stroke (NIH/NINDS)
R01 NS092695
United States
Citation
Journal: Mol Cell / Year: 2019 Title: Structures of the Rhodopsin-Transducin Complex: Insights into G-Protein Activation. Authors: Yang Gao / Hongli Hu / Sekar Ramachandran / Jon W Erickson / Richard A Cerione / Georgios Skiniotis / Abstract: Rhodopsin (Rho), a prototypical G-protein-coupled receptor (GPCR) in vertebrate vision, activates the G-protein transducin (G) by catalyzing GDP-GTP exchange on its α subunit (Gα). To elucidate the ...Rhodopsin (Rho), a prototypical G-protein-coupled receptor (GPCR) in vertebrate vision, activates the G-protein transducin (G) by catalyzing GDP-GTP exchange on its α subunit (Gα). To elucidate the determinants of G coupling and activation, we obtained cryo-EM structures of a fully functional, light-activated Rho-G complex in the presence and absence of a G-protein-stabilizing nanobody. The structures illustrate how G overcomes its low basal activity by engaging activated Rho in a conformation distinct from other GPCR-G-protein complexes. Moreover, the nanobody-free structures reveal native conformations of G-protein components and capture three distinct conformers showing the Gα helical domain (αHD) contacting the Gβγ subunits. These findings uncover the molecular underpinnings of G-protein activation by visual rhodopsin and shed new light on the role played by Gβγ during receptor-catalyzed nucleotide exchange.
History
Deposition
May 14, 2019
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Header (metadata) release
Jun 19, 2019
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Map release
Jul 24, 2019
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Update
Dec 4, 2019
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Current status
Dec 4, 2019
Processing site: RCSB / Status: Released
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Structure visualization
Movie
Surface view with section colored by density value
Cryogen name: ETHANE / Chamber humidity: 100 % / Instrument: FEI VITROBOT MARK III
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Electron microscopy
Microscope
FEI TITAN KRIOS
Image recording
Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Number real images: 12616 / Average exposure time: 8.0 sec. / Average electron dose: 50.0 e/Å2
Electron beam
Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron optics
C2 aperture diameter: 50.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
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Image processing
Particle selection
Number selected: 5872324
CTF correction
Software - Name: Gctf (ver. 1.06)
Final reconstruction
Resolution.type: BY AUTHOR / Resolution: 3.9 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cisTEM / Number images used: 250451
Initial angle assignment
Type: PROJECTION MATCHING
Final angle assignment
Type: MAXIMUM LIKELIHOOD
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Atomic model buiding 1
Refinement
Protocol: BACKBONE TRACE
Output model
PDB-6oy9: Structure of the Rhodopsin-Transducin Complex
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