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Title:negative stain single-particle reconstruction of conformation XXII of the Ltn1 E3 ubiquitin Ligase
Authors:Lyumkis D, Doamekpor SK, Bengtson MH, Lee JW, Toro TB, Petroski MD, Lima CD, Potter CS, Carragher B, Joazeiro CAP
Sample:snapshot XXII of Ltn1 conformer
Method:Single particle reconstruction (56.2 angstroms resolution)
Related entries in frame:EMD-2248, EMD-2249, EMD-2250, EMD-2251, EMD-2252, EMD-2253, EMD-2254, EMD-2255, EMD-2256, EMD-2257, EMD-2258, EMD-2259, EMD-2260, EMD-2261, EMD-2262, EMD-2263, EMD-2264, EMD-2265, EMD-2266, EMD-2267, EMD-2268
Red flagLatest update:2013-07-03
Sample
Sample name: snapshot XXII of Ltn1 conformer
Experimental molecular weight of the sample: 180
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
1proteinLtn1180180monomertrueSaccharomyces cerevisiae
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
8.00.01 mg/mL190 mM NaCl, 20 mM Tris, 1mM BME3 microliters of sample at a concentration of 0.01 mg/mL was applied to a C-flat grid with 2-micron-diameter holes overlaid by thin 1.5 nm continuous carbon. The specimen was stained with 2% uranyl formate 3 times, then let air-dry.c-flat grids overlaid with thin (~1.5 nm) carbon
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 F20120 kVFLOOD BEAMBRIGHT FIELD2 mm1200 nm2600 nm5000050000FIELD 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
single-tilt room temperatureSIDE ENTRY, EUCENTRIC-55° eV298 K K K mrad15 e/Å223-MAR-2011
Processing
Protocol:random-conical tilt reconstruction
Software:Spider, Appion
CTF correction:each micrograph
Number of particles:222
Number of class averages:1
Imposed symmetry:C1
Resolution by author:56.2 Å
Resolution method:FSC 0.5
Processing details:Random-conical tilt reconstruction.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
489 μm/pixel
Publicly available raw data:http://maskiton.scripps.edu
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails