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Electron tomography of isometrically contracting insect flight muscle
by electron tomography
#1: Solid view (volume rendering), , Made by UCSF CHIMERA
#2: Close-up, Solid view (volume rendering), , Made by UCSF CHIMERA
#3: Simplified surface model, Made by Jmol
 Entry | |
| Summary | |
| Database / ID | EM DATA BANK (EMDB) / 1561 |
|---|---|
| Title | Electron tomography of isometrically contracting insect flight muscle |
| Map | This is a dual axis tomogram of quick frozen, freeze This is a dual axis tomogram of quick frozen, freeze substituted, plastic embedded, thin sectioned insect flight muscle from Lethocerus sp. |
| Sample | Isometrically contracting asynchronous insect flight muscle |
| Keywords |  insect, muscle, myosin, troponin, tropomyosin, actin, light chains, thin filament, thick filament, electron microscopy, image processing, isometric contraction, freezing, freeze substitution, microtomy, multivariate data analysis |
| Authors | Wu S, Liu J, Reedy MC, Tregear RT, Winkler H, Franzini-Armstrong C, Sasaki H, Lucaveche C, Goldman YE, Reedy MK, Taylor KA |
| Date | Deposition: 2008-09-26, Header release: 2008-09-29, Map release: 2010-04-12, Last update: 2011-12-23 |
| EMDB Sites |  EMDB @PDBe (EU), EMDB @RCSB (USA) |
| Structure Visualization | |
| Movies |  Movie Page#1: Solid view (volume rendering), , Made by UCSF CHIMERA #2: Close-up, Solid view (volume rendering), , Made by UCSF CHIMERA #3: Simplified surface model, Made by Jmol |
| Supplemental images | |
| Structure viewers |  Yorodumi,  Launch PeppeR (About PeppeR), Volume viewer (RCSB, PDBe) |
| Related Structure Data | |
| Related Entries |
Cite: data citing same article |
 Article | |
| Citation - Primary | |
| Article | PLoS ONE, Vol. 5, Issue 9, Year 2010 |
|---|---|
| Title | Electron tomography of cryofixed, isometrically contracting insect flight muscle reveals novel actin-myosin interactions. |
| Authors | Shenping Wu, Jun Liu, Mary C Reedy, Richard T Tregear, Hanspeter Winkler, Clara Franzini-Armstrong, Hiroyuki Sasaki, Carmen Lucaveche, Yale E Goldman, Michael K Reedy, Kenneth A Taylor Institute of Molecular Biophysics, Florida State University, Tallahassee, Florida, United States of America. |
| Keywords | Actins (chemistry), Animals, Cryopreservation, Crystallography, X-Ray, Electron Microscope Tomography, Flight, Animal, Insect Proteins (chemistry), Insects (chemistry), Isometric Contraction, Models, Molecular, Muscles (chemistry), Myosins (chemistry, 3.6.4.1), Protein Binding, Protein Structure, Tertiary, Tissue Fixation |
| Links | DOI: 10.1371/journal.pone.0012643, PubMed: 20844746, PMC: PMC2936580 |
| Citation - #1 | |
| Article | J.STRUCT.BIOL., Vol. 168, Page 485-502, Year 2009 |
| Title | Methods for Identifying and Averaging Variable Molecular Conformations in Tomograms of Actively Contracting Insect Flight Muscle |
| Authors | Wu S, Liu J, Reedy MC, Tregear RT, Winkler H, Reedy MK, Taylor KA |
 Map | |||||||||||||||||||||
| File |  EMD-1561.map ( map file in CCP4 format, 602114 KB ) | ||||||||||||||||||||
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| Projections & Slices | Size of images:
Images are generated by Spider package. | ||||||||||||||||||||
| Density |
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| Data Type | Image stored as Reals | ||||||||||||||||||||
| Space Group Number | 1 | ||||||||||||||||||||
| Map Geometry |
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| Unit Cell | A: 9273.6 A, B: 11040 A, C: 483 A Alpha=beta=gamma: 90 degrees | ||||||||||||||||||||
| Pixel Spacing | X= Y= Z: 6.9 A | ||||||||||||||||||||
| CCP4 map header info | |||||||||||||||||||||
| Annotation Details | This is a dual axis tomogram of quick frozen, freeze This is a dual axis tomogram of quick frozen, freeze substituted, plastic embedded, thin sectioned insect flight muscle from Lethocerus sp. | ||||||||||||||||||||
 Supplement | |
| Images | |
| Images | |
|---|---|
 Sample | |
| Name | Isometrically contracting asynchronous insect flight muscle |
|---|---|
| Number of Components | 8 |
| Oligomeric 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 |
| Component #1: cellular-component - muscle | |
| Scientific name | myofibril |
| Common Name | muscle |
| Details | The sample was enblock stained using uranyl acetate and tannic acid, and post stained with lead citrate and potassium permanganate |
| Oligomeric Details | muscle fibril |
| Scientific Name of Species | Lethocerus indicus |
| Common Name of Species | Water bug |
| NCBI taxonomy | 212017 |
| Recombinant expression | No |
| Natural Source | Organ Or Tissue: asynchronous dorsal longitudinal flight muscle Organelle: myofibril Cell Location: myooplasm |
 Experiment | |
| Sample Preparation | |
| Staining | Freeze slammed fibers were freeze-substituted in acetone using a tannic aciduranyl acetate sequence, and ultimately embedded in Araldite-506 for thin-section electron microscopy. Ultrathin (25-30 nm) longitudinal sections were stained by permanganate-lead. |
|---|---|
| Buffer | Details: 20 mM MOPS buffer, 5 mM NaN3, and MgCl2, ATP, CaCl2, and EGTA in varying millimolar concentrations |
| Vitrification | |
| Method | smash against a liquid helium cooled Au-coated Cu mirror |
| Cryogen Name | HELIUM |
| Details | Vitrification instrument: modified Heuser Cryopress freezing head |
| Instrument | HOMEMADE PLUNGER |
| Temperature | 4.5 Kelvin |
| Imaging | |
| Microscope | FEI/PHILIPS CM300FEG/T |
| Electron Gun | |
| Electron Source | FIELD EMISSION GUN |
| Accelerating Voltage | 300 kV |
| Illumination Mode | FLOOD BEAM |
| Lens | |
| Nominal Cs | 2 mm |
| Imaging Mode | BRIGHT FIELD |
| Specimen Holder | |
| Holder | Gatan model 670 Ultrahigh tilt analytical holder |
| Model | OTHER |
| Tilt Angle | -72 degrees - 72 degrees |
| Camera | |
| Detector | Tietz TEM-CAM F224 |
| Image Acquisition | |
| Quant Bit Number | 16 |
| Details | Images recorded on a TVIPS Tem-Cam F224 2k x 2k CCD camera |
 Processing | |
| Method | electron tomography |
|---|---|
| 3D reconstruction | |
| Algorithm | marker free alignment |
| Software | PROTOMO, IMOD |
| Details | Two tilt series at 90 degrees to one another were first independently aligned using marker-free alignment and area matching. The two resulting tomograms were then merged by patch correlation and volume warp using IMOD. Tomogram computed using weighted back projection. |
| Resolution Method | FSC at 0.5 cut-off |
| Tomography | |
| Number of Sections | 70 |
| Download | |||
| Data from EMDB | |||
| Header (meta data in XML format) | emd-1561.xml (8.5 KB) | ||
|---|---|---|---|
| Map data | emd_1561.map.gz (532.4 MB) | ||
| Images |  1561.gif (189 KB) | ||
| FTP directory | ftp://ftp.pdbj.org/pub/emdb/structures/EMD-1561 | ||
| Movie files | |||
| movie #1 |
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| movie #2 |
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| movie #3 |
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