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Title:The eye lens chaperone alphaB-crystallin forms defined globular, 24meric assemblies
Authors:Peschek J, Braun N, Franzmann TM, Georgalis Y, Haslbeck M, Weinkauf S, Buchner J
Sample:Human alphaB crystallin
Method:Single particle reconstruction (20 angstroms resolution)
Red flagLatest update:2012-09-19
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Sample
Sample name: Human alphaB crystallin
Oligomeric state: 24-mer
Theoretical molecular weight of the sample: 0.485
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1proteinalphaB crystallin24mertrueHomo sapiens
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.40.05 mg/mL137mM NaCl, 2.7mM KCl, 12 mM PBSSamples were negatively stained for 30s using Ammonium Molybdate solution pH 5.5300 mesh carbon-coated copper grids
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
NONE% KNONE ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
JEOL 100CX100 kVFLOOD BEAMBRIGHT FIELD2.8 mm900 nm1200 nm50000OTHERKODAK SO-163 FILM mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
Side entryJEOL°° eV K K K mrad e/Å206-FEB-2008
Processing
Protocol:Cross-common lines
Software:Imagic
CTF correction:Phase flipping of each particle
Number of particles:2565
Number of class averages:40
Imposed symmetry:T
Resolution by author:20 Å
Resolution method:FSC 0.5
Processing details:Final maps were calculated from 40 class averages. After correction of contrast transfer function by phase flipping the reconstructions have been performed by iterative cycles of MRA, MSA, classification and angular reconstitution of the obtained class averages. See also publication Peschek et al., 2009. The eye lens chaperone alpha-crystallin forms defined globular assemblies. PNAS 106.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
111.69 μm/pixel16OTHER
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
1SHS rigid bodyChimeraRECIPROCALThe dimers were separately fitted by manual docking using program chimera. AS 14-103 were used