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    - PDB-2w4v: Isometrically contracting insect asynchronous flight muscle quick... -

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    Basic information

    Entry
    Database: PDB / ID: 2w4v
    TitleIsometrically contracting insect asynchronous flight muscle quick frozen after a quick release step
    DescriptorMYOSIN HEAVY CHAIN
    STRIATED MUSCLE
    MYOSIN REGULATORY LIGHT CHAIN
    STRIATED ADDUCTOR MUSCLE
    MYOSIN ESSENTIAL LIGHT CHAIN
    STRIATED ADDUCTOR MUSCLE
    KeywordsCONTRACTILE PROTEIN / TROPOMYOSIN / LIGHT CHAINS / ACTIN-BINDING / ISOMETRIC CONTRACTION / THIN FILAMENT / MOTOR PROTEIN / THICK FILAMENT / MUSCLE PROTEIN
    Specimen sourceArgopecten irradians / invertebrata / BAY SCALLOP
    MethodElectron microscopy (35 A resolution / Tomography / Freeze substituted insect flight muscle)
    AuthorsWu, S. / Liu, J. / Reedy, M.C. / Tregear, R.T. / Winkler, H. / Franzini-Armstrong, C. / Sasaki, H. / Lucaveche, C. / Goldman, Y.E. / Reedy, M.K. / Taylor, K.A.
    CitationPLoS ONE, 2012, 7, e39422-e39422

    primary. PLoS ONE, 2012, 7, e39422-e39422 StrPapers
    Structural changes in isometrically contracting insect flight muscle trapped following a mechanical perturbation.
    Shenping Wu / Jun Liu / Mary C Reedy / Robert J Perz-Edwards / Richard T Tregear / Hanspeter Winkler / Clara Franzini-Armstrong / Hiroyuki Sasaki / Carmen Lucaveche / Yale E Goldman / Michael K Reedy / Kenneth A Taylor

    #1. J.Struct.Biol., 2009 StrPapers
    Methods for Identifying and Averaging Variable Molecular Conformations in Tomograms of Actively Contracting Insect Flight Muscle.
    Wu, S. / Liu, J. / Reedy, M.C. / Winkler, H. / Reedy, M.K. / Taylor, K.A.

    DateDeposition: Dec 2, 2008 / Release: Sep 15, 2010 / Last modification: Jul 18, 2012

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    Assembly

    Deposited unit
    C: MYOSIN HEAVY CHAIN, STRIATED MUSCLE
    Y: MYOSIN REGULATORY LIGHT CHAIN, STRIATED ADDUCTOR MUSCLE
    Z: MYOSIN ESSENTIAL LIGHT CHAIN, STRIATED ADDUCTOR MUSCLE

    127 kDa, 3 molecules
    Theoretical massNumber of molelcules
    Total
    (without water)
    127,4723
    Polyers127,4723
    Non-polymers00
    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) / MYOSIN HEAVY CHAIN, STRIATED MUSCLE / MYOSIN S1 HEAVY CHAIN / MYOSIN S1 / Fragment: RESIDUES 5-835 / Source: ARGOPECTEN IRRADIANS (gene. exp.) / References: UniProt: P24733
    #2polypeptide(L) / MYOSIN REGULATORY LIGHT CHAIN, STRIATED ADDUCTOR MUSCLE / MYOSIN REGULATORY LIGHT CHAIN, RLC, STRIATED ADDUCTOR MUSCLE / MYOSIN S1, R-LC / Fragment: RESIDUES 16-151 / Source: ARGOPECTEN IRRADIANS (gene. exp.) / References: UniProt: P13543
    #3polypeptide(L) / MYOSIN ESSENTIAL LIGHT CHAIN, STRIATED ADDUCTOR MUSCLE / MYOSIN ESSENTIAL LIGHT CHAIN, ELC, STRIATED ADDUCTOR MUSCLE / MYOSIN S1, E-LC / Fragment: RESIDUES 5-155 / Source: ARGOPECTEN IRRADIANS (gene. exp.) / References: UniProt: P07291

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    Experimental details

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    Experiment

    ExperimentMethod: ELECTRON MICROSCOPY
    EM experimentReconstruction method: TOMOGRAPHY / Specimen type: FREEZE SUBSTITUTED INSECT FLIGHT MUSCLE

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    Sample preparation

    Assembly of specimenName: INSECT FIBRILLAR FLIGHT MUSCLE / Aggregation state: TISSUE
    Details: THIS SPECIMEN IS OBTAINED FROM A QUICK FROZEN, ISOMETRICALLY CONTRACTING ASYNCHRONOUS INSECT FLIGHT MUSCLE THAT HAS BEEN FREEZE SUBSTITUTED, PLASTIC EMBEDDED, AND THIN SECTIONED. THE FIBERS FOR THIS STUDY WERE SUBJECTED TO MECHANICAL TRANSIENTS. FOR THE QUICK RELEASE,THE FIBER WAS RELEASED 9 NM PER HALF-SARCOMERE IN 2 MS AND THE FREEZING IMPACT - OCC-6 MS LATER.
    Buffer solutionName: 20 MM MOPS BUFFER, 5 MM NAN3, AND MGCL2, ATP, CACL2, AND EGTA IN VARYING MILLIMOLAR CONCENTRATIONS
    Specimen supportDetails: CARBON
    VitrificationDetails: SMASH AGAINST A LIQUID HELIUM COOLED GOLD COATED COPPER MIRROR

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    Electron microscopy imaging

    MicroscopyMicroscope model: FEI/PHILIPS CM300FEG/T
    Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: LOW DOSE-FLOOD BEAM
    Electron lensMode: BRIGHT FIELD / Cs: 2 mm
    Specimen holderTilt angle max: 72 deg. / Tilt angle min: -72 deg.
    CameraType: TIETZ TEM-CAM F224
    Radiation wavelengthRelative weight: 1

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    Processing

    3D reconstructionDetails: NOTE THAT OUR LOWEST RESOLUTION DATA IS AT INVERSE 1 MICRON. NUMBER OF FOURIER COEFFICIENTS IS ALMOST A HALF MILLION. THESE COORDINATES WERE FITTED TO AVERAGED SUBVOLUMES OBTAINED FROM A DUAL AXIS TOMOGRAM. THE FITTING WAS DONE MANUALLY USING THE CRYSTALLOGRAPHIC MODEL FITTING PROGRAM O. THERE ARE 27 MODELS.
    Least-squares processHighest resolution: 35 A
    Refine hist #LASTHighest resolution: 35 A
    Number of atoms included #LASTProtein: 8501 / Nucleic acid: 0 / Ligand: 0 / Solvent: 0 / Total: 8501

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