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Title:3D reconstruction of frozen hydrated HIV-1 integrase dimer in complex with two Fabs.
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:HIV-1 integrase - Fab complex
Method:Single particle reconstruction (9.6 angstroms resolution)
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Sample
Sample name: HIV-1 integrase - Fab complex
Oligomeric state: 2 Fabs bind to one integrase dimer
Theoretical molecular weight of the sample: 0.16
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1proteinintegrase0.032dimertrueHuman immunodeficiency virus 1dimer, total molecular weight 65kDa
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
mg/mL200 mesh Quantifoil
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE100%100 KFEI VITROBOT msVitrification instrument: Vitrobot
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TECNAI F20200 kVFLOOD BEAMBRIGHT FIELD2.1 mm2000 nm5000 nm80000FIELD EMISSION GUNTVIPS TEMCAM-F816 (8k x 8k) mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
CT3500GATAN LIQUID NITROGEN eV K K K mrad30 e/Å201-FEB-2011
Processing
Protocol:Fourier space
Software:Frealign
CTF correction:Each particle
Number of particles:14000
Imposed symmetry:C2
Resolution by author:9.6 Å
Resolution method:FSC 0.143
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
1EXQ rigid bodyChimeraREALProtocol: rigid body
1M71 rigid bodyChimeraREALProtocol: rigid body