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Title:Negative stain EM structure of the HOPS tethering complex (conformation 1)
Authors:Broecker C, Kuhlee A, Gatsogiannis C, Balderhaar HJ, Hoenscher C, Engelbrecht-Vandre S, Ungermann C, Raunser S
Sample:vacuole protein sorting (HOPS) tethering complex containing Vps11, Vps16, Vps18, Vps33, Vp39, Vps41
Method:Single particle reconstruction (30.4 angstroms resolution)
Sample
Sample name: vacuole protein sorting (HOPS) tethering complex containing Vps11, Vps16, Vps18, Vps33, Vp39, Vps41
Oligomeric state: Monomer
Theoretical molecular weight of the sample: 0.7
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
1proteinvacuolar protein sorting-associated protein 41MonomerfalseSaccharomyces cerevisiaeP38959GO:0030897
2proteinvacuolar protein sorting-associated protein 39MonomerfalseSaccharomyces cerevisiaeQ07468GO:0030897
3proteinvacuolar protein sorting-associated protein 33MonomerfalseSaccharomyces cerevisiaeP20795GO:0030897
4proteinvacuolar protein sorting-associated protein 18MonomerfalseSaccharomyces cerevisiaeP27801GO:0030897
5proteinvacuolar protein sorting-associated protein 11MonomerfalseSaccharomyces cerevisiaeP12868GO:0030897
6proteinvacuolar protein sorting-associated protein 16MonomerfalseSaccharomyces cerevisiaeQ03308GO:0030897
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.5 mg/mL1 M NaCl, 50 mM Hepes/NaOH, 10% glycerolGrids with adsorbed Protein floated on 0.07% Uranyl Formate for 1-3h at 4C.glow discharged 400 mesh copper grids with thin carbon support
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
NONE% KNONE ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
JEOL 1400120 kVFLOOD BEAMBRIGHT FIELD3.4 mm1.3 nm1.7 nm50000LAB6KODAK SO-163 FILM mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
JEOL°° eV K K K mrad4.5 e/Å207-FEB-2011
Processing
Protocol:Projection matching
Software:SPARX
Number of particles:2979
Imposed symmetry:C1
Resolution by author:30.4 Å
Resolution method:FSC 0.5
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
92010.5 μm/pixel16OTHER
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails