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Title:Cryo-EM Structure of the Mycobacterial Fatty Acid Synthase
Authors:Boehringer D, Ban N, Leibundgut M
Sample:Mycobacterial Fatty Acid Synthase, FAS I
Method:Single particle reconstruction (7.5 angstroms resolution)
Red flagLatest update:2013-02-27
Sample
Sample name: Mycobacterial Fatty Acid Synthase, FAS I
Oligomeric state: hexamer
Experimental molecular weight of the sample: 2
Components:
ID Type Name Exp. MW (MDa) Theo. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1proteinMycobacterial Fatty Acid Synthase IhexamerfalseMycobacterium smegmatisGO:0005835IPR003965
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.21.65 mg/mL100mM potassium phosphate buffer pH 7.2, 165 mM NaCl, 2mM EDTA, 2mM DTTQuantifoil R2/1 200 mesh copper grids
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE% KHOMEMADE PLUNGER ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TITAN KRIOS300 kVFLOOD BEAMBRIGHT FIELD2.7 mm1500 nm5000 nm59000100000FIELD EMISSION GUNFEI FALCON I (4k x 4k) mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
FEI TITAN KRIOS AUTOGRID HOLDER eV79 K K K mrad20 e/Å2Data were collected using the automated image acquisition software FEI EPU.19-OCT-2012
Processing
Protocol:Common lines, projection matching
Software:Imagic-5, Spider
CTF correction:Each image
Number of particles:106884
Imposed symmetry:D3
Resolution by author:7.5 Å
Resolution method:FSC 0.5
Processing details:Fourier amplitudes of the reconstruction were enhanced using amplitudes from the x-ray structure of the S. cerevisiae FAS; subsequently, the map was filtered using a Butterworth low-pass filter in SPIDER.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
1556 μm/pixel16Data were collected using the automated image acquisition software FEI EPU.
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
2UV8 rigid bodyCorrelationChimeraREALProtocol: Domains were separately fitted as rigid bodies and manually adjusted in O. The model was minimized with PHENIX.PDBTOOLS. Domains were separately fitted as rigid bodies and manually adjusted in O.The model was minimized with PHENIX.PDBTOOLS.