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TitleAtomic model of the F420-reducing [NiFe] hydrogenase by electron cryo-microscopy using a direct electron detector.
Journal, issue, pagesElife, Vol. 3, Page e01963, Year 2014
Publish dateFeb 25, 2014
AuthorsMatteo Allegretti / Deryck J Mills / Greg McMullan / Werner Kühlbrandt / Janet Vonck /
PubMed AbstractThe introduction of direct electron detectors with higher detective quantum efficiency and fast read-out marks the beginning of a new era in electron cryo-microscopy. Using the FEI Falcon II direct ...The introduction of direct electron detectors with higher detective quantum efficiency and fast read-out marks the beginning of a new era in electron cryo-microscopy. Using the FEI Falcon II direct electron detector in video mode, we have reconstructed a map at 3.36 Å resolution of the 1.2 MDa F420-reducing hydrogenase (Frh) from methanogenic archaea from only 320,000 asymmetric units. Videos frames were aligned by a combination of image and particle alignment procedures to overcome the effects of beam-induced motion. The reconstructed density map shows all secondary structure as well as clear side chain densities for most residues. The full coordination of all cofactors in the electron transfer chain (a [NiFe] center, four [4Fe4S] clusters and an FAD) is clearly visible along with a well-defined substrate access channel. From the rigidity of the complex we conclude that catalysis is diffusion-limited and does not depend on protein flexibility or conformational changes. DOI: http://dx.doi.org/10.7554/eLife.01963.001.
External linksElife / PubMed:24569482 / PubMed Central
MethodsEM (single particle)
Resolution3.36 Å
Structure data

2513

4ci0

EMDB-2513: Electron cryo-microscopy of F420-reducing [NiFe] hydrogenase Frh
PDB-4ci0: Electron cryo-microscopy of F420-reducing NiFe hydrogenase Frh
Method: EM (single particle) / Resolution: 3.36 Å

Chemicals

ChemComp-FE:
Unknown entry

ChemComp-NI:
NICKEL (II) ION

ChemComp-FE2:
Unknown entry

ChemComp-SF4:
IRON/SULFUR CLUSTER

ChemComp-ZN:
Unknown entry

ChemComp-FAD:
FLAVIN-ADENINE DINUCLEOTIDE / FAD*YM

Source
  • methanothermobacter marburgensis (archaea)
KeywordsOXIDOREDUCTASE / FLAVOPROTEIN / ELECTRON TRANSFER / FERREDOXIN

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