top_cornerHomeEMD-2096  Contact EMDataBank 
Image unavailable
Title:Cryo-EM structure of the F420-reducing [NiFe]-hydrogenase from a methanogenic archaeon at near-atomic resolution
Authors:Mills DJ, Vitt S, Strauss M, Shima S, Vonck J
Sample:F420-reducing [NiFe] hydrogenase Frh
Method:Single particle reconstruction (4.5 angstroms resolution)
Related entries in frame:EMD-2097
Red flagLatest update:2013-07-03
Sample
Sample name: F420-reducing [NiFe] hydrogenase Frh
Oligomeric state: dodecamer
Theoretical molecular weight of the sample: 1.2
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1proteinF420-reducing [NiFe] hydrogenase1.2dodecamerfalseMethanothermobacter marburgensisIPR017682, IPR017679, IPR017681The Frh complex is a tetrahedral dodecamer of an FrhA, FrhB, FrhG heterotrimer
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.60.7 mg/mL50mM Tris-HCl, 0.025mM FADfreshly glow discharged Quantifoil holey grid (1 micrometer holes)
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE70%103 KFEI VITROBOT MARK Iblotting 2.5 seconds before plunging ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI POLARA 300200 kVFLOOD BEAMBRIGHT FIELD2 mm1690 nm2800 nm5900061400FIELD EMISSION GUNKODAK SO-163 FILM mmObjective lens astigmatism was corrected at 120,000 times magnification

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
OTHER°° eV79 K77 K80 K mrad15 e/Å226-AUG-2010
Processing
Software:EMAN2
CTF correction:each micrograph
Number of particles:84000
Number of class averages:2134
Resolution by author:4.5 Å
Resolution method:feature-based
Processing details:EMAN2.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
101 μm/pixel8ZEISS SCAI
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
2WPN rigid bodychimera, cootREALProtocol: A rigid body fit was done in Chimera and the model manually rebuilt in Coot.