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Title:Helical reconstruction of HMPV matrix protein-lipid filaments
Authors:Leyrat C, Renner M, Harlos K, Huiskonen JT, Grimes JM
Sample:Human metapneumovirus matrix protein M bound to DOPC
Method:Helical reconstruction (28 angstroms resolution)
Red flagLatest update:2016-02-17
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Sample
Sample name: Human metapneumovirus matrix protein M bound to DOPC
Oligomeric state: helical
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1proteinhuman metapneumovirus matrix proteindimertrueHuman metapneumovirusQ91F56GO:0019068, GO:0019031IPR005056matrix protein was mixed with DOPC at a final concentration of 0.4 mM
2ligand1,2-Dioleoyl-sn-glycero-3-phosphocholinefalsesynthetic construct
Experiment
Specimen state: Filament
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.50.2 mg/mL20 mM Tris, 650 mM NaCl, 1M NDSB-201Grids with adsorbed protein-lipid mixtures floated on 2% w/v uranyl acetate for 30 seconds300 mesh copper grid with formvar carbon film
Helical parameters:
Phi incrementAzimuthal (Z) increment
56.5°5.16 Å
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
NONE% KNONE ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TECNAI F30200 kVFLOOD BEAMBRIGHT FIELD mm720 nm820 nm3900048387FIELD EMISSION GUNGATAN ULTRASCAN 4000 (4k x 4k) mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
SIDE ENTRY, EUCENTRIC°° eV295 K K K mrad e/Å216-JUL-2013
Processing
Protocol:Helical reconstruction
Software:Burnham-Brandeis, Helical, Package
CTF correction:Each micrograph
Resolution by author:28 Å
Resolution method:Last visible layer line
Processing details:Final map was filtered to 28 angstrom resolution. The reconstruction was calculated using Burnham-Brandeis Helical Package.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
115 μm/pixel12
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails