Electron cryomicroscopy structure of N-ethyl maleimide sensitive factor at 11 A resolution.

REL PDBe PDBe PDBe 2003-10-10 2003-10-10 2003-10-10 2012-10-24 Electron cryomicroscopy structure of N-ethyl maleimide sensitive factor at 11 A resolution. Furst J Sutton RB Chen J Brunger AT Grigorieff N Furst J Sutton RB Chen J Brunger AT Grigorieff N Electron cryomicroscopy structure of N-ethyl maleimide sensitive factor at 11 A resolution. EMBO J. 22 4365 4374 2003 12941689 doi:10.1093/emboj/cdg420 NSF alpha-SNAP SNARE 20S NSF alpha-SNAP SNARE 20S

The sample was monodisperse. Some domains were disordered.

3 0.68 gel permeation chromatography N-ethyl Maleimide Sensitive Factor Cricetulus griseus Chinese Hamster 0.0828

6 subunits

6 hexamer true Escherichia coli Soluble NSF Attachment Protein Bos taurus Cattle 0.0334 true Soluble NSF Attachment Protein Receptor unidentified Chinese Hamster 0.0823 true singleParticle particle 0.2 7.4

20mM HEPES, pH 7.4, 300 mM NACl, 2 mM MgCl2, 5mM b-mercaptoethanol, 0.01/0.1 mM ADP/ATP nucleotide

NEGATIVE

cryo (no stain)

holey carbon film

ETHANE 80 85 HOMEMADE PLUNGER

Vitrification instrument: Technion plunger

FEI TECNAI 20 FLOOD BEAM BRIGHT FIELD FIELD EMISSION GUN 200 2.0 2.0 4.0 62000.0 GATAN LIQUID NITROGEN 85 KODAK SO-163 FILM ZEISS SCAI 7 10

3 x 3 averaging, final step size = 27 microns

1.4 8. Eucentric 0 0

manual particle selection

FREALIGN

C6 OTHER 11.0 FSC 0.5 CUT-OFF IMAGIC, FREALIGN

Final map was calculated from three data sets

31592 emd_1059.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 90 90 90 1 1 1 90 90 90 315 315 315 90 90 90 X Y Z -0.681735 1.45625 0.0257073 0.145834 3.5 3.5 3.5 0.144 3D density map of NSF?alpha-SNAP?SNARE (20S)

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