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Title:Structure of a stalled transfer intermediate of Sm proteins from the assembly chaperone pICln to the SMN-complex
Authors:Chari A, Grimm C, Fischer U, Stark H
Sample:8S complex: a stalled Sm protein transfer intermediate from the assembly chaperone pICln to the SMN-complex
Method:Single particle reconstruction (20 angstroms resolution)
Red flagLatest update:2013-03-06
Sample
Sample name: 8S complex: a stalled Sm protein transfer intermediate from the assembly chaperone pICln to the SMN-complex
Oligomeric state: One heterooctamer composed of pICln, SmD1, SmD2, SmE, SmF, SmG, SMN and Gemin2
Experimental molecular weight of the sample: 0.125
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
1protein8S complex0.1250.125monomertrueHomo sapiens
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.510 mg/mL20 mM HEPES, 150 mM NaCl, 5 mM DTTGrids were floated on 2 % Uranyl Formate solution for 1 minute.Custom made holey grid with thin carbon support.
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
NONE% KNONE msnegative stain
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI/PHILIPS CM200FEG160 kVSPOT SCANBRIGHT FIELD2 mm nm nm109000FIELD EMISSION GUNGENERIC CCD mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
SIDE ENTRY, EUCENTRIC°° eV K K K mrad20 e/Å212-JAN-2012
Processing
Protocol:exact filter backprojection
Software:Imagic
CTF correction:Each particle
Number of particles:10000
Imposed symmetry:C1
Resolution by author:20 Å
Resolution method:FSC 0.5
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
48 μm/pixel16
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
4F77 rigid bodyAmiraREALProtocol: Rigid Body