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Title:Octameric structure of IMPDH from Pseudomonas aeruginosa in its APO form
Authors:Labesse G, Vaupre L, Salard-Arnaud I, Alexandre T, Lai Kee Him J, Raynal B, Bron P, Munier-Lehmann H
Sample:IMPDH apo filtered at 15.5 A resolution from Pseudomonas aeruginosa
Method:Single particle reconstruction (15.5 angstroms resolution)
Red flagLatest update:2014-03-26
Sample
Sample name: IMPDH apo filtered at 15.5 A resolution from Pseudomonas aeruginosa
Oligomeric state: octameric
Theoretical molecular weight of the sample: 0.43
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
1proteinInosine-5'-monophosphate dehydrogenase0.43octamertruePseudomonas aeruginosa
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
8.01 mg/mL20 mM potassium phosphate pH 8 and 0.1 M KClQuantifoil R2.2, glow discharged
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE98%102.15 KGATAN CRYOPLUNGE 3Blot for 1 second ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
JEOL 2200FS200 kVFLOOD BEAMBRIGHT FIELD2.0 mm1400 nm2500 nm100000130137FIELD EMISSION GUNGENERIC GATAN (4k x 4k) mmObjective lens astigmatism was corrected at 200,000 times magnification

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
GATAN LIQUID NITROGEN°°JEOL0-20 eV95.15 K93.15 K96.15 K mrad18 e/Å2Weak beam illumination01-OCT-2011
Processing
Software:IMAGIC, FREALIGN
CTF correction:Each particle
Number of particles:10625
Imposed symmetry:D4
Resolution by author:15.5 Å
Resolution method:FSC 0.5
Processing details:The particles were selected using an semi-automatic selection program (boxer).
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
74 μm/pixelImages recorded with the 4K X 4K slow-scan CCD camera (Gatan inc.), which explains why the nominal magnification is 100000 and 130137 when calibrated on the camera.
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails