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Title:Negative stain EM structure of the PTC3 holotoxin complex (TcdA1, TcdB2, TccC3) in pore state
Authors:Gatsogiannis C, Lang A E, Meusch D, Pfaumann V, Hofnagel O, Benz R, Aktories K, Raunser S
Sample:PTC3 (TcdA1, TcdB2, TccC3)
Method:Single particle reconstruction (40 angstroms resolution)
Red flagLatest update:2013-04-03
Sample
Sample name: PTC3 (TcdA1, TcdB2, TccC3)
Oligomeric state: TcdB2/TccC3 bind to one homopentamer of TcdA1
Experimental molecular weight of the sample: 1.68
Components:
ID Type Name Exp. MW (MDa) Theo. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1proteinTcdA10.2830.283truePhotorhabdus luminescensQ9RN43
2proteinTcdB20.1660.166truePhotorhabdus luminescensQ7N7Y2
3proteinTccC30.1090.109truePhotorhabdus luminescensQ7N7Y4
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
11 mg/mL50 mM CAPS, 100 mM NaCl, 0.05% Tween-20, 5% glycerolGrids with adsorbed Protein floated on 0.07% Uranyl Formate for 45 sec.glow discharged 400 mesh copper grids with thin carbon support
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
NONE% KNONE ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
JEOL 1400120 kVFLOOD BEAMBRIGHT FIELD3.4 mm1.2 nm2 nm5000067210LAB6TVIPS TEMCAM-F416 (4k x 4k) mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
JEOL°° eV K K K mrad e/Å217-JAN-2012
Processing
Protocol:Projection matching
Software:Sparx
Number of particles:317
Imposed symmetry:C5
Resolution by author:40 Å
Resolution method:FSC 0.5
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
104 μm/pixel
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails