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Title:Cryo-electron microscopy of phirsl1 jumbo phage
Authors:Effantin G, Hamasaki R, Kawasaki T, Bacia M, Moriscot C, Weissenhorn W, Yamada T, Schoehn G
Sample:phirsl1 helical portion of the tail
Method:Helical reconstruction (9.6 angstroms resolution)
Sample
Sample name: phirsl1 helical portion of the tail
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
1proteinTail tube protein0.018falseRalstonia phage RSL1UNP B2ZY00
2proteinTail sheath protein0.068falseRalstonia phage RSL1IPR007067UNP B2ZXZ8
Experiment
Specimen state: Filament
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.5 mg/mL50 mM Tris-HCl [pH 7.5], 100 mM NaCl, 10 mM MgSO4Four microliters of the bacteriophage sample (~0.1 mg/ml) were loaded between the mica-carbon interface. The sample was stained using 2% ammonium molybdate pH 7.5 and air-dried.
Helical parameters:
HandPhi incrementAzimuthal (Z) incrementRotational symmetry
RIGHT HANDED22.04°37.9 Åc6
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE100% KFEI VITROBOT MARK IV ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI POLARA 300300 kVSPOT SCANBRIGHT FIELD2.3 mm1355 nm5295 nm31000FIELD EMISSION GUNKODAK SO-163 FILM mmObjective lens astigmatism was corrected at 59000

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
GATAN LIQUID NITROGEN°° eV K K K mrad20 e/Å225-FEB-2011
Processing
Protocol:projection matching
Software:SPIDER, IHRSR
CTF correction:each particle
Resolution by author:9.6 Å
Resolution method:FSC 0.5
Processing details:Projection matching was done with SPIDER. IHRSR was used for helical parameters search and application.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
527 μm/pixel8ZEISS SCAI
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails