Journal: Elife / Year: 2018 Title: Architecture of the human mTORC2 core complex. Authors: Edward Stuttfeld / Christopher Hs Aylett / Stefan Imseng / Daniel Boehringer / Alain Scaiola / Evelyn Sauer / Michael N Hall / Timm Maier / Nenad Ban / Abstract: The mammalian target of rapamycin (mTOR) is a key protein kinase controlling cellular metabolism and growth. It is part of the two structurally and functionally distinct multiprotein complexes mTORC1 ...The mammalian target of rapamycin (mTOR) is a key protein kinase controlling cellular metabolism and growth. It is part of the two structurally and functionally distinct multiprotein complexes mTORC1 and mTORC2. Dysregulation of mTOR occurs in diabetes, cancer and neurological disease. We report the architecture of human mTORC2 at intermediate resolution, revealing a conserved binding site for accessory proteins on mTOR and explaining the structural basis for the rapamycin insensitivity of the complex.
History
Deposition
Oct 15, 2017
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Header (metadata) release
Dec 20, 2017
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Map release
Feb 28, 2018
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Update
Feb 28, 2018
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Current status
Feb 28, 2018
Processing site: PDBe / Status: Released
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Structure visualization
Movie
Surface view with section colored by density value
Cryogen name: ETHANE-PROPANE / Chamber humidity: 95 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV / Details: Two seconds blotting.
Details
Sample was prepared by a modified GRAFIX protocol described in the paper, and was extremely dilute. Particles were adhered to a thin carbon film to obtain reasonable coverage.
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Electron microscopy
Microscope
FEI TITAN KRIOS
Temperature
Min: 100.0 K / Max: 100.0 K
Specialist optics
Energy filter - Name: GIF Quantum SE
Image recording
Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Digitization - Dimensions - Width: 3838 pixel / Digitization - Dimensions - Height: 3710 pixel / Digitization - Sampling interval: 5.0 µm / Digitization - Frames/image: 1-40 / Number grids imaged: 1 / Number real images: 3997 / Average exposure time: 20.0 sec. / Average electron dose: 80.0 e/Å2 / Details: Dose weighting applied
Electron beam
Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Software - Name: CTFFIND (ver. 4) / Details: Performed in Relion
Startup model
Type of model: OTHER Details: Continuation from EMDB entry EMD-3927 after subtraction of mTOR-mLST8 and focused refinement of the accessory factor density
Final reconstruction
Number classes used: 1 / Applied symmetry - Point group: C1 (asymmetric) / Algorithm: FOURIER SPACE / Resolution.type: BY AUTHOR / Resolution: 6.2 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 2.0) Details: Continuation from EMDB entry EMD-3927 after subtraction of mTOR-mLST8 and focused refinement of the accessory factor density Number images used: 83362
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