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Title:Molecular architecture and conformational flexibility of human RNA polymerase II.
Authors:S.A. Kostek, P. Grob, S. De Carlo, J. S. Lipscomb, F. Garczarek and E. Nogales
Sample:Human RNA Polymerase II
Aggregation state:Single particle (20 angstroms resolution)
Red flagLatest update:2011-05-26
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bullet_white.gif  Summary
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bullet.png Experimental details
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Sample
Sample name: Human RNA Polymerase II
Oligomeric state: 12 subunit complex
Theoretical molecular weight of the sample: 0.52
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism GO identifier InterPro identifier Virus identifier Details
1proteinRpb10.22falseHomo sapiens
2proteinRpb20.13falseHomo sapiens
3proteinRpb30.03falseHomo sapiens
4proteinRpb40.02falseHomo sapiens
5proteinRpb50.02falseHomo sapiens
6proteinRpb60.01falseHomo sapiens
7proteinRpb70.02falseHomo sapiens
8proteinRpb80.02falseHomo sapiens
9proteinRpb90.01falseHomo sapiens
10proteinRpb100.01falseHomo sapiens
11proteinRpb110.01falseHomo sapiens
12proteinRpb120.01falseHomo sapiens
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
7.90.01 mg/mL200mM (NH4)2SO4, 25 mM Hepes, 0.2mM EDTA, 0.05% NP-40cryonegative staining in a saturated solution of ammonium molybdate neutralized to a pH of 7.2. with 10 N NaOH400 mesh copper grid
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE%90 Kstained in ammonium molybdate before plunging ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI/PHILIPS CM200FEG200 kVFLOOD BEAMBRIGHT FIELD2.2 mm1200 nm3600 nm5000050280FIELD EMISSION GUNKodak SO163 film mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
side entryGATAN LIQUID NITROGEN°° eV90 K K K mrad17 e/Å2
Processing
Software:FREALIGN
CTF correction:each micrograph
Resolution by author:20 Å
Resolution method:FSC at 0.143 cut-off
Processing details:conformation 1 and 2 were obtained after 3D variance analysis and targeted classification
Unit cell:
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
196.3 μm/pixel1.014linkOTHER
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
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