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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 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
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
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
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
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
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
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
2QXV rigid bodyChimeraREALProtocol: local rigid-body fitting algorithm
2YB8 rigid bodyChimeraREALProtocol: local rigid-body fitting algorithm