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Title:3D reconstruction of negatively stained PCSK9 in complex with a Fab
Authors:Wu S, Avila-Sakar A, Kim J, Booth DS, Greenberg CH, Rossi A, Liao M, Alian A, Griner SL, Juge N, Mergel CM, Chaparro-Riggers J, Strop P, Tampe R, Edwards RH, Stroud RM, Craik CS, Cheng Y
Sample:human PCSK9 in complex with a Fab
Method:Single particle reconstruction (25 angstroms resolution)
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Sample
Sample name: human PCSK9 in complex with a Fab
Oligomeric state: monomer
Theoretical molecular weight of the sample: 0.12
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1proteinPCSK90.12monomertrueHomo sapiens
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
mg/mLuranyl formate200 mesh copper grid
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 20120 kVFLOOD BEAMBRIGHT FIELD2.0 mm1.5 nm1.5 nm5000050000LAB6GATAN ULTRASCAN 4000 (4k x 4k) mmeach particle

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
single tiltSIDE ENTRY, EUCENTRIC60° eV K K K mrad30 e/Å2
Processing
Protocol:Fourier space
Software:Frealign
CTF correction:each particle
Number of particles:3876
Imposed symmetry:C1
Resolution by author:25 Å
Resolution method:FSC 0.5
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
2PMW REAL