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Title:Substrate Recruitment Pathways in the Yeast Exosome by Electron Microscopy
Authors:Liu J-J, Bratkowski MA, Liu XQ, Niu C-Y, Ke AL, Wang H-W
Sample:Rrp44-Exosome incubated with RNA36
Method:Single particle reconstruction (23 angstroms resolution)
Red flagLatest update:2014-01-15
Sample
Sample name: Rrp44-Exosome incubated with RNA36
Experimental molecular weight of the sample: 0.350
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
1proteinRrp440.11falseSaccharomyces cerevisiae
2proteinRrp430.04falseSaccharomyces cerevisiae
3proteinRrp40.04falseSaccharomyces cerevisiae
4proteinCsl40.03falseSaccharomyces cerevisiae
5proteinRrp450.03falseSaccharomyces cerevisiae
6proteinRrp46-TAP0.03trueSaccharomyces cerevisiae
7proteinRrp410.03falseSaccharomyces cerevisiae
8proteinRrp420.03falseSaccharomyces cerevisiae
9proteinMtr30.03falseSaccharomyces cerevisiae
10proteinRrp400.03falseSaccharomyces cerevisiae
11nucleic-acidRNA360.01truesynthetic construct
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
8.01 mg/mL150mM NaCl, 50mM Tris-HCL,1mM DTTAll samples were diluted at a final concentration of ~80 nM of the exosome in the non-digestive buffer and negatively stained in 2% (w/v) uranyl acetate solution following the standard deep stain procedure on holey-carbon coated EM copper grids covered with a thin layer of continuous carbon300 mesh continus carbon
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
NONE% KNONE ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TECNAI F20200 kVFLOOD BEAMBRIGHT FIELD mm nm nmFIELD EMISSION GUNGENERIC GATAN (4k x 4k) mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
OTHEROTHER°°FEI eV K K K mrad30 e/Å210-OCT-2012
Processing
Software:Spider
Number of particles:23711
Imposed symmetry:C1
Resolution by author:23 Å
Resolution method:FSC 0.5
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
300 μm/pixel
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
4IFD rigid bodyChimeraREAL