Journal: Nat Struct Mol Biol / Year: 2016 Title: Cryo-electron microscopy structure of the TRPV2 ion channel. Authors: Lejla Zubcevic / Mark A Herzik / Ben C Chung / Zhirui Liu / Gabriel C Lander / Seok-Yong Lee / Abstract: Transient receptor potential vanilloid (TRPV) cation channels are polymodal sensors involved in a variety of physiological processes. TRPV2, a member of the TRPV family, is regulated by temperature, ...Transient receptor potential vanilloid (TRPV) cation channels are polymodal sensors involved in a variety of physiological processes. TRPV2, a member of the TRPV family, is regulated by temperature, by ligands, such as probenecid and cannabinoids, and by lipids. TRPV2 has been implicated in many biological functions, including somatosensation, osmosensation and innate immunity. Here we present the atomic model of rabbit TRPV2 in its putative desensitized state, as determined by cryo-EM at a nominal resolution of ∼4 Å. In the TRPV2 structure, the transmembrane segment 6 (S6), which is involved in gate opening, adopts a conformation different from the one observed in TRPV1. Structural comparisons of TRPV1 and TRPV2 indicate that a rotation of the ankyrin-repeat domain is coupled to pore opening via the TRP domain, and this pore opening can be modulated by rearrangements in the secondary structure of S6.
History
Deposition
Sep 4, 2015
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Header (metadata) release
Oct 14, 2015
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Map release
Jan 20, 2016
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Update
Feb 17, 2016
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Current status
Feb 17, 2016
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 % / Chamber temperature: 120 K / Instrument: GATAN CRYOPLUNGE 3 / Method: Blot for 2.5 seconds before plunging.
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Electron microscopy
Microscope
FEI TITAN KRIOS
Temperature
Min: 77 K / Max: 79 K / Average: 78 K
Alignment procedure
Legacy - Astigmatism: Objective astigmatism was corrected at 22500 times magnification using Thon rings visualized with a K2 camera.
Details
Data was acquired using Leginon and collected in K2 super-resolution mode.
Date
May 12, 2015
Image recording
Category: CCD / Film or detector model: GATAN K2 (4k x 4k) / Number real images: 747 / Average electron dose: 57 e/Å2 / Details: 50 frames, 200 ms per frame
Electron beam
Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Pre-processing was performed in the Appion pipeline using a template for particle selection, CTFFIND3 for CTF estimation, MSA/MRA for 2D classification, and RELION for 3D classification and refinement.
CTF correction
Details: whole micrograph
Final reconstruction
Algorithm: OTHER / Resolution.type: BY AUTHOR / Resolution: 3.8 Å / Resolution method: OTHER / Software - Name: Appion, CTFFIND3, FindEM, EMAN, RELION Details: Per-frame dose weighting was performed using the RELION particle polishing method. Number images used: 43879
FSC plot (resolution estimation)
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