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Title:Antibody Recognition of the Pandemic H1N1 Influenza Hemagglutinin Receptor Binding Site
Authors:Hong M, Lee PS, Hoffman RMB, Zhu X, Krause JC, Laursen NS, Yoon S, Song L, Tussey L, Crowe Jr JE, Ward AB, Wilson IA
Sample:Influenza hemagglutinin (A/Singapore/6/1986, H1N1) bound to neutralizing antibody (Fab 5J8)
Method:Single particle reconstruction (22 angstroms resolution)
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Sample
Sample name: Influenza hemagglutinin (A/Singapore/6/1986, H1N1) bound to neutralizing antibody (Fab 5J8)
Oligomeric state: One HA trimer bound to three Fabs
Theoretical molecular weight of the sample: 0.289
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1protein5J8 Fab fragmentmonomertrueHomo sapiens
2proteinInfluenza A/Singapore/6/1986 (H1N1) hemagglutinintrimertrueInfluenza A virus
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.40.02 mg/mL150 mM NaCl, 50 mM Tristwo cycles of "Nano-W" (2% methylamine tungstate) applied for 20 seconds400 mesh copper grids coated in nitrocellulose and thin carbon, glow discharged in argon/oxygen
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 12120 kVFLOOD BEAMBRIGHT FIELD mm1300 nm1300 nm5200052000LAB6Tietz TemCam-416 CMOS mmObjective lens astigmatism corrected at 100,000 times magnification.

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
SIDE ENTRY, EUCENTRIC0°55° eV298 K K K mrad e/Å214-FEB-2013
Processing
Protocol:Projection matching initiated with low-pass filtered hemagglutinin
Software:Appion, Spider, Xmipp, Eman1
Number of particles:5106
Imposed symmetry:C3
Resolution by author:22 Å
Resolution method:FSC at 0.5 cut-off (FSC calculated with Eman1's eotest procedure.)
Processing details:Particles were selected using automatic (difference-of-Gaussians) picking followed by reference-free classification to eliminate noisy picks or non-target aggregation states.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
306 μm/pixelTilt series varying from 0-55 degrees, alternating low and high tilts on similarly stained regions
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails