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Title:Structure of ABCA4, the photoreceptor-specific ATP-binding cassette transporter, in complex with AMPPNP
Authors:Tsybovsky Y, Orban T, Molday RS, Taylor D, Palczewski K
Sample:ABCA4 purified from bovine rod outer segments and supplemented with AMPPNP, a non-hydrolyzable ATP analogue
Method:Single particle reconstruction (18.9 angstroms resolution)
Red flagLatest update:2013-05-22
Sample
Sample name: ABCA4 purified from bovine rod outer segments and supplemented with AMPPNP, a non-hydrolyzable ATP analogue
Oligomeric state: monomer
Experimental molecular weight of the sample: 0.4
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
1proteinABCA40.40.36monomerfalseBos taurusThe total molecular weight includes estimated MW of Amphipol 8-35 bound to the protein
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.50.1 mg/mL20 mM BTP, pH 7.5, 150 mM NaCl, 10% glycerol, 1 mM DTT, 1 mM AMPPNPGrids with adsorbed protein were stained with 1% w/v uranyl acetate for 10 secondsGlow discharged Cu/Pd 400 mesh 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
FEI TECNAI F20200 kVOTHERBRIGHT FIELD mm200 nm1500 nm5000070373FIELD EMISSION GUN mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
OTHER45° eV K K K mrad e/Å221-APR-2012
Processing
Protocol:reference-based projection alignment
Software:Spider
CTF correction:Each defocus group
Number of particles:38090
Imposed symmetry:C1
Resolution by author:18.9 Å
Resolution method:FSC 0.5
Processing details:The particles were selected manually.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
200 μm/pixel
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails