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Title:Cryo-EM structure of the F420-reducing [NiFe]-hydrogenase from a methanogenic archaeon with bound substrate
Authors:Mills DJ, Vitt S, Strauss M, Shima S, Vonck J
Sample:F420-reducing [NiFe] hydrogenase Frh with bound substrate
Method:Single particle reconstruction (4.5 angstroms resolution)
Related entries in frame:EMD-2096
Red flagLatest update:2013-04-03
Sample name: F420-reducing [NiFe] hydrogenase Frh with bound substrate
Oligomeric state: dodecamer
Theoretical molecular weight of the sample: 1.2
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. The substrate F420 is bound to FrhB
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)
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE70%103 KFEI VITROBOT MARK Iblotting 2.5 seconds before plunging ms
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI POLARA 300200 kVFLOOD BEAMBRIGHT FIELD2 mm1650 nm3550 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/Å211-FEB-2011
CTF correction:each micrograph
Number of particles:97000
Number of class averages:2134
Imposed symmetry:T
Resolution by author:4.5 Å
Resolution method:feature-based
Processing details:EMAN2.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
63 μm/pixel8ZEISS SCAI
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.