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Title:Structure of TcdA1 pore (TcdA1 reconstituted in liposomes)
Authors:Gatsogiannis C, Lang A E, Meusch D, Pfaumann V, Hofnagel O, Benz R, Aktories K, Raunser S
Sample:TcdA1
Method:Single particle reconstruction (19.9 angstroms resolution)
Red flagLatest update:2013-04-03
Sample
Sample name: TcdA1
Oligomeric state: Pentamer
Experimental molecular weight of the sample: 1.41
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 luminescensQ9RN43Protein reconstituted in liposomes
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
11 mg/mL50 mM CAPS pH 11, 100 mM NaCl, 5 % glycerolC-Flat 1.2/1.3-4C copper 400 mesh, with additional thin carbon support
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE90%100 KGATAN CRYOPLUNGE 3Blot for 1 sec before plunging ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
JEOL 3200FSC200 kVFLOOD BEAMBRIGHT FIELD4.1 mm2.9 nm5 nm4000085460FIELD EMISSION GUNTVIPS TEMCAM-F816 (8k x 8k) mmObjective lens astigmatism was corrected at 120,000 times magnification

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
JEOL 3200FSC CRYOHOLDER°°in-column Omega filter0-15 eV K K K mrad20 e/Å218-MAR-2012
Processing
Protocol:Projection matching
Software:Sparx
CTF correction:Each Particle
Number of particles:5549
Imposed symmetry:C5
Resolution by author:19.9 Å
Resolution method:FSC 0.5
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
145 μm/pixel
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails