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- EMDB-7029: CryoEM map from poorly ordered myosin thick filaments isolated fr... -

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Basic information

Entry
Database: EMDB / ID: 7029
TitleCryoEM map from poorly ordered myosin thick filaments isolated from asynchronous flight muscle of the large waterbug Lethocerus indicus
Map dataPrimary map
SampleLethocerus flight muscle myosin filament:
SourceLethocerus indicus (insect)
Methodsingle particle reconstruction / cryo EM / 6.4 Å resolution
AuthorsTaylor KA / Taylor D / Hu Z / Edwards RJ
CitationJournal: J. Struct. Biol. / Year: 2017
Title: Coupling between myosin head conformation and the thick filament backbone structure.
Authors: Zhongjun Hu / Dianne W Taylor / Robert J Edwards / Kenneth A Taylor
Abstract: The recent high-resolution structure of the thick filament from Lethocerus asynchronous flight muscle shows aspects of thick filament structure never before revealed that may shed some light on how ...The recent high-resolution structure of the thick filament from Lethocerus asynchronous flight muscle shows aspects of thick filament structure never before revealed that may shed some light on how striated muscles function. The phenomenon of stretch activation underlies the function of asynchronous flight muscle. It is most highly developed in flight muscle, but is also observed in other striated muscles such as cardiac muscle. Although stretch activation is likely to be complex, involving more than a single structural aspect of striated muscle, the thick filament itself, would be a prime site for regulatory function because it must bear all of the tension produced by both its associated myosin motors and any externally applied force. Here we show the first structural evidence that the arrangement of myosin heads within the interacting heads motif is coupled to the structure of the thick filament backbone. We find that a change in helical angle of 0.16° disorders the blocked head preferentially within the Lethocerus interacting heads motif. This observation suggests a mechanism for how tension affects the dynamics of the myosin heads leading to a detailed hypothesis for stretch activation and shortening deactivation, in which the blocked head preferentially binds the thin filament followed by the free head when force production occurs.
DateDeposition: Sep 15, 2017 / Header (metadata) release: Oct 11, 2017 / Map release: Oct 11, 2017 / Last update: Feb 14, 2018

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.012
  • Imaged by UCSF Chimera
  • Download
  • Surface view colored by cylindrical radius
  • Surface level: 0.012
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

Downloads & links

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Map

Fileemd_7029.map.gz (map file in CCP4 format, 322487 KB)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
432 pix
1.22 Å/pix.
= 528.336 Å
432 pix
1.22 Å/pix.
= 528.336 Å
432 pix
1.22 Å/pix.
= 528.336 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.223 Å
Density
Contour Level:0.032 (by author), 0.012 (movie #1):
Minimum - Maximum-0.056497645 - 0.18061732
Average (Standard dev.)0.00056291226 (0.009138624)
Details

EMDB XML:

Space Group Number1
Map Geometry
Axis orderXYZ
Dimensions432432432
Origin000
Limit431431431
Spacing432432432
CellA=B=C: 1 Å
α=β=γ: 90 deg.

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.2231.2231.223
M x/y/z432432432
origin x/y/z0.0000.0000.000
length x/y/z528.336528.336528.336
α/β/γ90.00090.00090.000
start NX/NY/NZ
NX/NY/NZ
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS432432432
D min/max/mean-0.0560.1810.001

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Supplemental data

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Sample components

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Entire Lethocerus flight muscle myosin filament

EntireName: Lethocerus flight muscle myosin filament / Number of components: 1

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Component #1: protein, Lethocerus flight muscle myosin filament

ProteinName: Lethocerus flight muscle myosin filament / Recombinant expression: No
SourceSpecies: Lethocerus indicus (insect)
Source (natural)Organelle: myofibril / Organ or tissue: striated muscle

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Experimental details

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Sample preparation

SpecimenSpecimen state: filament / Method: cryo EM
Sample solutionpH: 6.8
Support filmGatan Solarus 950.M plasma cleaner
VitrificationInstrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Temperature: 295 K / Humidity: 100 %

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Electron microscopy imaging

Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
ImagingMicroscope: FEI TITAN KRIOS
Details: All data collection details may be found in Z. Hu, D. W. Taylor, M. K. Reedy, R. J. Edwards, K. A. Taylor, Structure of myosin filaments from relaxed Lethocerus flight muscle by cryo-EM at 6 Angstrom resolution. Sci. Adv. 2, e1600058 (2016). Can also be found in the Specimen section of EMD-3301
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 65 e/Å2 / Illumination mode: FLOOD BEAM
LensImaging mode: BRIGHT FIELD / Defocus: 1500 - 3000 nm
Specimen HolderModel: FEI TITAN KRIOS AUTOGRID HOLDER
CameraDetector: OTHER

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Image acquisition

Image acquisitionDetails: All specimen and sample preparation details may be found in Z. Hu, D. W. Taylor, M. K. Reedy, R. J. Edwards, K. A. Taylor, Structure of myosin filaments from relaxed Lethocerus flight muscle by cryo-EM at 6 Angstrom resolution. Sci. Adv. 2, e1600058 (2016). Can also be found in the Specimen section of EMD-3301

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Image processing

ProcessingMethod: single particle reconstruction / Applied symmetry: C4 (4 fold cyclic) / Number of projections: 50000
Details: All specimen and sample preparation details may be found in Z. Hu, D. W. Taylor, M. K. Reedy, R. J. Edwards, K. A. Taylor, Structure of myosin filaments from relaxed Lethocerus flight muscle by cryo-EM at 6 Angstrom resolution. Sci. Adv. 2, e1600058 (2016). Can also be found in the Specimen section of EMD-3301
3D reconstructionSoftware: RELION
CTF correction: All specimen and sample preparation details may be found in Z. Hu, D. W. Taylor, M. K. Reedy, R. J. Edwards, K. A. Taylor, Structure of myosin filaments from relaxed Lethocerus flight muscle by cryo-EM at 6 Angstrom resolution. Sci. Adv. 2, e1600058 (2016). Can also be found in the Specimen section of EMD-3301
Resolution: 6.4 Å / Resolution method: FSC 0.143 CUT-OFF
FSC plot
(resolution estimation)

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