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Title:Negative Staining Structure of Human Polycomb Repressive Complex 2 bound to the co-factor AEBP2
Authors:Ciferri C, Lander GC, Maiolica A, Herzog F, Aebersold R, Nogales E
Sample:Negative Staining Reconstruction of Polycomb repressive Complex 2 bound to the co-factor AEBP2
Method:Single particle reconstruction (21 angstroms resolution)
Sample
Sample name: Negative Staining Reconstruction of Polycomb repressive Complex 2 bound to the co-factor AEBP2
Oligomeric state: Pentameric Complex
Theoretical molecular weight of the sample: 265
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
1proteinRbAp48monomertrueHomo sapiensGO:0035098IPR001214
2proteinEzh2monomertrueHomo sapiensGO:0035098IPR001214
3proteinSuz12monomertrueHomo sapiensGO:0035098IPR001214
4proteinAEBP2monomertrueHomo sapiensGO:0035098IPR001214
5proteinEEDmonomertrueHomo sapiensGO:0035098IPR001214
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.60.1 mg/mL25 mM Hepes pH 7.5, 138 mM NaCl, 10% Glycerol, 0.05% NP40, 1 mM Tris(2-carboxyethyl) phosphine hydrochloride [TCEP]2% Uranyl Formate400 mesh copper carbon support
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 20120 kVFLOOD BEAMDARK FIELD2.2 mm600 nm1300 nm8000080000FIELD EMISSION GUNGENERIC GATAN (4k x 4k) mmObjective lens astigmatism was corrected at 150,000 times magnification

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
OTHERFEI eV298 K K K mrad20 e/Å211-APR-2011
Processing
Protocol:Projection matching from RCT model
Software:Appion, Leginon, Spider, Imagic, EMAN2, SPARX
CTF correction:whole micrograph
Number of particles:39527
Number of class averages:1000
Imposed symmetry:C1
Resolution by author:21 Å
Resolution method:FSC 0.5
Processing details:Material and Methods section of Paper.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
500 μm/pixel
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
2QXV rigid bodyChimeraREALProtocol: local rigid-body fitting algorithm
2YB8 rigid bodyChimeraREALProtocol: local rigid-body fitting algorithm