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Title:The structure and super-organization of acetylcholine receptor-rapsyn complexes; class C
Authors:Zuber B, Unwin N
Sample:Postsynaptic membrane isolated from Torpedo marmorata electric organ
Method:Subtomogram averaging (42 angstroms resolution)
Red flagLatest update:2013-07-03
Sample
Sample name: Postsynaptic membrane isolated from Torpedo marmorata electric organ
Oligomeric state: acetylcholine receptors bound to two pairs of rapsyns
Theoretical molecular weight of the sample: 0.474
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
1proteinnicotinic acetylcholine receptor0.2900.280falseTorpedo marmorataIPR002394
2proteinRapsyn0.0430.046falseTorpedo marmorataGO:0007268, GO:0005515, GO:0008270, GO:0033130, GO:0005856, GO:0030054, GO:0045202, GO:0045211IPR001237, IPR019568, IPR011990, IPR013026, IPR013083, IPR001841, IPR019734, IPR001440, IPR018293the number of copies is suggested by the volume of the rapsyn densities
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.4 mg/mL400 mM NaCl, 20 mM phosphate buffer, leupeptin 0.3 mg/l, pepstatin 1 mg/lunstained300 mesh copper grid with lacy carbon support, glow discharge in amylamine atmosphere
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE90%78 KHOMEMADE PLUNGERblot from the carbon side ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI POLARA 300300 kVFLOOD BEAMBRIGHT FIELD2 mm3000 nm6000 nm6900080213FIELD EMISSION GUNGATAN ULTRASCAN 4000 (4k x 4k) mmobjective lens astigmatism was corrected at 69,000 times magnification

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
GATAN LIQUID NITROGEN-70°70°Gatan0-20 eV85 K80 K95 K mrad50 e/Å2dual tilt axis tomography25-JUL-2008
Processing
Protocol:Simultaneous Iterative Reconstruction Technique (SIRT)
Software:IMOD, PRIISM/IVE
Number of subtomogram averaged:3564
Number of class averages:5
Imposed symmetry:C1
Resolution by author:42 Å
Resolution method:FSC 0.5
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
14215 μm/pixel16dual axis tilt series
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
2BG9 rigid bodyChimeraREAL