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Title:Electron cryo-microscopy of a cross-beta amyloid fibril polymorph
Authors:Fitzpatrick AWP, Debelouchina GT, Bayro MJ, Clare DK, Caporini MA, Bajaj VS, Jaroniec CP, Wang L, Ladizhansky V, Muller S, MacPhee CE, Waudby CA, Mott HR, De-Simone A, Knowles TPJ, Saibil HR, Vendruscolo M, Orlova EV, Griffin RG, Dobson CM
Sample:Doublet cross-beta amyloid fibril polymorph
Method:Single particle reconstruction (12.7 angstroms resolution)
Red flagLatest update:2013-11-06
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Sample name: Doublet cross-beta amyloid fibril polymorph
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1proteinTransthyretin(105-115)falseRattus norvegicus
Specimen state: Filament
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
21 mg/mL10% acetonitrile/water solutionholey carbon films (R2/2, QuantifoilMicro Tools GmbH, Jena, Germany)
Helical parameters:
HandPhi incrementAzimuthal (Z) increment
LEFT HANDED0.85°4.67 Å
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE95%99 KHOMEMADE PLUNGERFibrils were applied to holey carbon films that were immediately plunge-frozen at liquid nitrogen temperature. ms
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TECNAI F20200 kVFLOOD BEAMBRIGHT FIELD mm900 nm3000 nm4000040000FIELD EMISSION GUNKODAK SO-163 FILM mmObjective lens astigmatism was corrected at 100,000 times magnification.

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
Nitrogen cooledGATAN LIQUID NITROGEN eV99 K99 K100 K mrad e/Å203-MAR-2006
Protocol:Common crossovers
CTF correction:CTFFIND3
Number of particles:82
Number of class averages:18
Imposed symmetry:C1
Resolution by author:12.7 Å
Resolution method:FSC 0.5
Processing details:The particles were selected using an automatic selection program.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
250 μm/pixelZEISS SCAI
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails