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    - PDB-3zuh: Negative stain EM Map of the AAA protein CbbX, a red-type Rubisco... -

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    Basic information

    Entry
    Database: PDB / ID: 3zuh
    TitleNegative stain EM Map of the AAA protein CbbX, a red-type Rubisco activase from R. sphaeroides
    DescriptorPROTEIN CBBX
    KeywordsATP BINDING PROTEIN / AAA+ PROTEIN
    Specimen sourceRhodobacter sphaeroides / archaea
    MethodElectron microscopy (21 A resolution / Single particle / Negative stain)
    AuthorsMueller-Cajar, O. / Stotz, M. / Wendler, P. / Hartl, F.U. / Bracher, A. / Hayer-Hartl, M.
    CitationNature, 2011, 479, 194-199

    Nature, 2011, 479, 194-199 StrPapers
    Structure and function of the AAA+ protein CbbX, a red-type Rubisco activase.
    Oliver Mueller-Cajar / Mathias Stotz / Petra Wendler / F Ulrich Hartl / Andreas Bracher / Manajit Hayer-Hartl

    DateDeposition: Jul 19, 2011 / Release: Nov 9, 2011 / Last modification: Aug 27, 2014

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    Assembly

    Deposited unit
    A: PROTEIN CBBX
    B: PROTEIN CBBX
    C: PROTEIN CBBX
    D: PROTEIN CBBX
    E: PROTEIN CBBX
    F: PROTEIN CBBX
    hetero molecules

    200 kDa, 18 molecules
    Theoretical massNumber of molelcules
    Total
    (without water)
    200,10318
    Polyers195,6796
    Non-polymers4,42412
    Water0

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    #1idetical with deposited unit / defined by author&software (PISA) / Symmetry operations: (identity)x1
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    Components

    #1polypeptide(L) / PROTEIN CBBX / THE PROTEIN IS BOUND TO RIBULOSE-1,5-BISPHOSPHATE / RUBISCO ACTIVASE CBBX / Source: RHODOBACTER SPHAEROIDES (gene. exp.) / References: UniProt: P95648
    #2ChemComp-RUB / SUGAR (RIBULOSE-1,5-DIPHOSPHATE)
    #3ChemComp-ADP / ADENOSINE-5'-DIPHOSPHATE / ADP *YM

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    Experimental details

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    Experiment

    ExperimentMethod: ELECTRON MICROSCOPY
    EM experimentReconstruction method: SINGLE PARTICLE / Specimen type: NEGATIVE STAIN

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    Sample preparation

    Assembly of specimenName: PROTEIN CBBX / Aggregation state: PARTICLE
    Buffer solutionName: 20 MM TRIS PH 8.0, 50 MM NACL, 5MM MGCL2, 1MM RIBULOSE-1,5- BISPHOSPHATE, 1MM ATP/ATPGAMMAS
    Sample preparationpH: 8 / Sample conc.: 0.07 mg/ml
    Specimen supportDetails: CARBON

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    Electron microscopy imaging

    MicroscopyMicroscope model: FEI TECNAI 12 / Date: Nov 19, 2010
    Electron gunElectron source: LAB6 / Accelerating voltage: 120 kV / Electron dose: 20 e/A2 / Illumination mode: LOW DOSE
    Electron lensMode: BRIGHT FIELD / Nominal magnification: 90600 X / Calibrated magnification: 90600 X / Nominal defocus max: 1800 nm / Nominal defocus min: 260 nm / Cs: 2 mm
    CameraType: EAGLE 2K CCD
    EM image scansNumber digital images: 12
    Radiation wavelengthRelative weight: 1

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    Processing

    Image selectionSoftware name: MRC, IMAGIC, SPIDER / Number of particles: 245
    EM single particle entitySymmetry type: MIXED SYMMETRY
    3D reconstructionMethod: ANGULAR RECONSTITUTION / Resolution: 21 A / Nominal pixel size: 3.308 A/pix / Actual pixel size: 3.308 A/pix / CTF correction method: PHASE FLIPPING, EACH PARTICLE
    Details: MODULE CONSISTING OF CHAIN A (204-296) AND CHAIN B (8-203) AS FOUND IN PDB 3SYL WAS MODELLED ONTO D2 P97 OF PDB 3CF3. SUBMISSION BASED ON EXPERIMENTAL DATA FROM EMDB EMD-1932.
    Atomic model buildingMethod: RIGID BODY / Ref protocol: MANUAL / Ref space: REAL
    Least-squares processHighest resolution: 21 A
    Refine hist #LASTHighest resolution: 21 A
    Number of atoms included #LASTProtein: 13614 / Nucleic acid: 0 / Ligand: 270 / Solvent: 0 / Total: 13884

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