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Title:Structure of the capsaicin receptor, TRPV1, determined by single particle electron cryo-microscopy
Authors:Liao M, Cao E, Julius D, Cheng Y
Sample:Rat TRPV1
Method:Single particle reconstruction (3.275 angstroms resolution)
Red flagLatest update:2015-10-14
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Sample
Sample name: Rat TRPV1
Oligomeric state: tetramer
Theoretical molecular weight of the sample: 0.3
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1proteinTRPV10.3TetramertrueRattus norvegicusO35433Functional minimal construct containing residues 110-603 and 627-764.
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.40.3 mg/mL150 mM NaCl, 20 mM HEPES, 2 mM TCEP400 mesh Quantifoil grid
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE90%120 KFEI VITROBOT MARK IIIBlot for 6 sec ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI POLARA 300300 kVFLOOD BEAMBRIGHT FIELD2.0 mm1500 nm3000 nm3100031000FIELD EMISSION GUNGATAN K2 (4k x 4k) mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
Cooled to Liquid Nitrogen temperatureOTHER°° eV K K K mrad21 e/Å2Gatan K2 Summit in super-resolution counting mode. Motion correction as described in Li et al. (2013) Nature Methods.01-JAN-2013
Processing
Protocol:Maximum likelihood
Software:RELION
CTF correction:Each particle
Number of particles:35645
Imposed symmetry:C4
Resolution by author:3.275 Å
Resolution method:Gold standard FSC at 0.143 cut-off
Processing details:3D classification, refinement, and reconstruction were performed using RELION.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
946 μm/pixelEvery image is the average of 30 frames recorded using the K2 Summit. The final reconstruction was calculated from images averaged from frames #3-#16.
Publicly available raw data:http://dx.doi.org/10.6019/EMPIAR-10005
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails