Journal: J Struct Biol / Year: 2007 Title: Octomeric pyruvate-ferredoxin oxidoreductase from Desulfovibrio vulgaris. Authors: Florian Garczarek / Ming Dong / Dieter Typke / H Ewa Witkowska / Terry C Hazen / Eva Nogales / Mark D Biggin / Robert M Glaeser / Abstract: Pyruvate-ferredoxin oxidoreductatse (PFOR) carries out the central step in oxidative decarboxylation of pyruvate to acetyl-CoA. We have purified this enzyme from Desulfovibrio vulgaris Hildenborough ...Pyruvate-ferredoxin oxidoreductatse (PFOR) carries out the central step in oxidative decarboxylation of pyruvate to acetyl-CoA. We have purified this enzyme from Desulfovibrio vulgaris Hildenborough (DvH) as part of a systematic characterization of as many multiprotein complexes as possible for this organism, and the three-dimensional structure of this enzyme has been determined by a combination of electron microscopy (EM), single particle image analysis, homology modeling and computational molecular docking. Our results show that the 1MDa DvH PFOR complex is a homo-octomer, or more precisely, a tetramer of the dimeric form of the related enzyme found in Desulfovibrio africanus (Da), with which it shares a sequence identity of 69%. Our homology model of the DvH PFOR dimer is based on the Da PFOR X-ray structure. Docking of this model into our 17A resolution EM-reconstruction of negatively stained DvH PFOR octomers strongly suggests that the difference in oligomerization state for the two species is due to the insertion of a single valine residue (Val383) within a surface loop of the DvH enzyme. This study demonstrates that the strategy of intermediate resolution EM reconstruction coupled to homology modeling and docking can be powerful enough to infer the functionality of single amino acid residues.
History
Deposition
Dec 27, 2006
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Header (metadata) release
Feb 5, 2007
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Map release
Jun 21, 2007
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Update
May 26, 2011
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Current status
May 26, 2011
Processing site: PDBe / Status: Released
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Structure visualization
Movie
Surface view with section colored by density value
Specimen holder: side entry / Specimen holder model: OTHER
Image recording
Category: FILM / Film or detector model: KODAK SO-163 FILM / Digitization - Scanner: OTHER / Digitization - Sampling interval: 12.7 µm / Number real images: 19
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Image processing
CTF correction
Details: defocus groups
Final reconstruction
Applied symmetry - Point group: D4 (2x4 fold dihedral) / Resolution.type: BY AUTHOR / Resolution: 17.0 Å / Resolution method: FSC 0.5 CUT-OFF / Software - Name: EMAN, SPIDER / Number images used: 12402
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