0000000000587831

AUTHOR

Dirk Scheffler

showing 5 related works from this author

Formation, TEM study and 3D reconstruction of the human erythrocyte peroxiredoxin-2 dodecahedral higher-order assembly.

2004

The production of a higher-order assembly of peroxiredoxin-2 (Prx-2) from human erythrocytes has been achieved during specimen preparation on holey carbon support films, in the presence of ammonium molybdate and polyethylene glycol. TEM study suggested that this assembly is a regular dodecahedron, containing 12 Prx-2 decamers (Mr 2.62 MDa, external diameter approximately 20 nm). This interpretation has been supported by production of a approximately 1.6 nm 3D reconstruction from the negative stain TEM data, with automated docking of the available X-ray data of the Prx-2 decamer. Comparison with other known protein dodecahedral and viral icosahedral structures indicates that this arrangement…

Models MolecularMaterials scienceErythrocytesIcosahedral symmetryMacromolecular SubstancesMacromolecular SubstancesGeneral Physics and AstronomyCell BiologyPeroxiredoxin 2Polyethylene glycolPeroxiredoxinsNegative stainDodecahedronCrystallographychemistry.chemical_compoundProtein structurechemistryMicroscopy Electron TransmissionPeroxidasesStructural BiologyImage Processing Computer-AssistedHumansGeneral Materials ScienceProtein Structure QuaternaryMacromoleculeMicron (Oxford, England : 1993)
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Interaction of the Vibrio cholerae cytolysin (VCC) with cholesterol, some cholesterol esters, and cholesterol derivatives: a TEM study.

2002

The Vibrio cholerae cytolysin (VCC) 63-kDa monomer has been shown to interact in aqueous suspension with cholesterol microcystals to produce a ring/pore-like heptameric oligomer approximately 8 nm in outer diameter. Transmission electron microscopy data were produced from cholesterol samples adsorbed to carbon support films, spread across the holes of holey carbon films, and negatively stained with ammonium molybdate. The VCC oligomers initially attach to the edge of the stacked cholesterol bilayers and with increasing time cover the two planar surfaces. VCC oligomers are also released into solution, with some tendency to cluster, possibly via the hydrophobic membrane-spanning domain. At th…

ErgosterolLiposomeCytotoxinsTemperatureOligomerNegative stainProtein Structure TertiaryCrystallographychemistry.chemical_compoundMicroscopy ElectronMonomerCholesterolchemistryPulmonary surfactantModels ChemicalStructural BiologySide chainImage Processing Computer-Assistedlipids (amino acids peptides and proteins)CytolysinCholesterol EstersVibrio choleraeFluorescent DyesJournal of structural biology
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Comparison of the decameric structure of peroxiredoxin-II by transmission electron microscopy and X-ray crystallography

2001

Abstract The decameric human erythrocyte protein torin is identical to the thiol-specific antioxidant protein-II (TSA-II), also termed peroxiredoxin-II (Prx-II). Single particle analysis from electron micrographs of Prx-II molecules homogeneously orientated across holes in the presence of a thin film of ammonium molybdate and trehalose has facilitated the production of a ≥20 A 3-D reconstruction by angular reconstitution that emphasises the D5 symmetry of the ring-like decamer. The X-ray structure for Prx-II was fitted into the transmission electron microscopic reconstruction by molecular replacement. The surface-rendered transmission electron microscopy (TEM) reconstruction correlates well…

Models MolecularMolybdenumErythrocytesSurface PropertiesCryo-electron microscopyChemistryResolution (electron density)BiophysicsTrehaloseSingle particle analysisPeroxiredoxinsCrystallography X-RayBiochemistryNegative stainMicroscopy ElectronCrystallographyPeroxidasesElectron tomographyStructural BiologyTransmission electron microscopyHumansEnergy filtered transmission electron microscopyOrthorhombic crystal systemMolecular BiologyBiochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology
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Haliotis tuberculata hemocyanin (HtH): analysis of oligomeric stability of HtH1 and HtH2, and comparison with keyhole limpet hemocyanin KLH1 and KLH2

2000

The multimeric/higher oligomeric states of the two isoforms of Haliotis tuberculata hemocyanin (HtH1 and HtH2) have been assessed by transmission electron microscopy (TEM) of negatively stained specimens, for comparison with previously published structural data from keyhole limpet hemocyanin (KLH1 and KLH2) [see Harris, J.R., Gebauer, W., Guderian, F.U., Markl, J., 1997a. Keyhole limpet hemocyanin (KLH), I: Reassociation from Immucothel followed by separation of KLH1 and KLH2. Micron, 28, 31-41; Harris, J.R., Gebauer, W., Söhngen, S.M., Nermut, M.V., Markl, J., 1997b. Keyhole limpet hemocyanin (KLH). II: Characteristic reassociation properties of purified KLH1 and KLH2. Micron, 28, 43-56; H…

medicine.medical_treatmentProtein subunitMagnesium ChlorideGeneral Physics and Astronomychemistry.chemical_elementCalciumOligomerCalcium Chloridechemistry.chemical_compoundStructural BiologymedicineAnimalsProtein IsoformsGeneral Materials ScienceMagnesium ionbiologyMagnesiumHemocyaninCell BiologyNegative stainMicroscopy ElectronCrystallographychemistryMolluscaHemocyaninsbiology.proteinBiophysicsKeyhole limpet hemocyaninMicron
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Routine preparation of air-dried negatively stained and unstained specimens on holey carbon support films: a review of applications.

2002

Several representative examples are given of the successful application of negative staining across the holes of holey carbon support films using 5% (w/v) ammonium molybdate solution containing trehalose. The inclusion of 0.1% (w/v) trehalose is considered to be most satisfactory, although good data have also been obtained in the presence of 0.01 and 1.0% (w/v) trehalose. The examples given fall into the following groups: protein molecules in the absence of polyethylene glycol (PEG), protein molecules in the presence of PEG (Mr 1000), lipoproteins, lipids and membranes, filaments and tubules, viruses in the absence of PEG, viruses in the presence of PEG, aqueous polymer solutions, and final…

Materials sciencePolymersLipoproteinsGeneral Physics and AstronomyPolyethylene glycolPolyethylene Glycolschemistry.chemical_compoundStructural BiologyPEG ratioAnimalsHumansGeneral Materials ScienceAmmoniumAmmonium molybdateOrganellesAqueous solutionStaining and LabelingHistological TechniquesProteinsTrehaloseCell BiologyNegative stainTrehaloseLipidsCarbonCrystallographyMicroscopy ElectronMembranechemistryHemocyaninsVirusesNuclear chemistryMicron (Oxford, England : 1993)
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