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Title:Cryo-EM structure of short shafted adenovirus type 5 complexed with factor X
Authors:Doronin K, Flatt JW, Di Paolo NC, Khare R, Kalyuzhniy O, Acchione M, Sumida JP, Ohto U, Shimizu T, Akashi-Takamura S, Miyake K, MacDonald JW, Bammler TK, Beyer RP, Farin FM, Stewart PL, Shayakhmetov DM
Sample:Complex of Ad5-short-fiber with factor X
Method:Single particle reconstruction (11.1 angstroms resolution)
Red flagLatest update:2012-11-21
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Sample
Sample name: Complex of Ad5-short-fiber with factor X
Oligomeric state: 240 factor X molecules bind to one Ad virion
Theoretical molecular weight of the sample: 164
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1virusHuman adenovirus 5Human adenovirus 5
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.60.6 mg/mL50mM Tris, 150mM NaCl, 2mM CaCl2, 2mM MgCl2Quantifoil R2/4 holey carbon grids, glow discharged
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE30%90 KHOMEMADE PLUNGERBlot for 6 seconds before plunging ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI POLARA 300300 kVFLOOD BEAMBRIGHT FIELD2.26 mm500 nm2100 nm310000400000FIELD EMISSION GUNGATAN ULTRASCAN 4000 (4k x 4k) mmObjective lens astigmatism was corrected at 300,000 times magnification

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
OTHER°° eV90 K K K0 mrad20 e/Å2Low dose12-NOV-2010
Processing
Protocol:Fourier methods
Software:Frealign
CTF correction:Each particle
Number of particles:520
Imposed symmetry:I
Resolution by author:11.1 Å
Resolution method:FSC 0.5
Processing details:The particles were selected with an in-house script and processed using Frealign.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
110115 μm/pixel16OTHER
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails