REL PDBe PDBe PDBe 2003-02-04 2003-07-09 2003-07-09 2011-05-26 The cyanide degrading nitrilase from Pseudomonas stutzeri AK61 is a two-fold symmetric, 14-subunit spiral. Sewell BT Berman MN Meyers PR Jandhyala D Benedik MJ Sewell BT Berman MN Meyers PR Jandhyala D Benedik MJ The cyanide degrading nitrilase from Pseudomonas stutzeri AK61 is a two-fold symmetric, 14-subunit spiral. STRUCTURE 11 1413 1422 2003 14604531 Cyanide dihydratase from Pseudomonas stutzeri Cyanide dihydratase from Pseudomonas stutzeri
The sample eluted as a single symmetric peak on gel filtration chromatography. Three bands were seen on SDS-PAGE presumably indicating some proteolysis
14 identical subunits 1 0.5 0.532 gel filtration chromatography, sequencing and counting subunits. cyanide dihydratase Pseudomonas stutzeri AK61 bacteria bacteria cytoplasm 0.038 0.038
The protein elutes as a sharp symmetric peak on Sephacryl 300 HR chromatography and particles appear homogeneously sized by negative stain EM
14 14mer true Escherichia coli pET26b GO:0000257 IPR000132 singleParticle particle 0.3 8.0
10 mM triethanolamine, 50 mM NaCl
 NEGATIVE
A carbon-coated copper grid was placed on a droplet of this enzyme preparation for 10 minutes, blotted, and then stained with 2% uranyl acetate for 3 minutes. The grid was then blotted and air-dried.
carbon film on 200 mesh copper grid
 NONE JEOL 2000EXII FLOOD BEAM BRIGHT FIELD TUNGSTEN HAIRPIN 120 5.6 0.1 0.7 50000.0 OTHER
Additional details about microscope model:JEOL 1200EXII
2001-03-01 KODAK SO-163 FILM 20 15 16. Eucentric
particles were picked manually
none
 C2 OTHER 25.0 FSC 0.5 CUT-OFF SPIDER
Initially no symmetry was assumed but a two fold axis became apparent after 47 cycles. The orientation of the twofold axis was determined and the model was rotated so that the twofold axis was parallel to the x-axis. In 17 subsequent cycles of refinement the twofold symmetry was imposed by rotating about the x-axis and adding the structure thus formed to the original.
7008
generated at 15 degree intervals by the SPIDER VO EA function.
 84 emd_1050.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 80 80 80 0 0 0 80 80 80 320 320 320 90 90 90 X Y Z -0.329862 0.6125 -0.00721576 0.0641706 4 4 4 0.153
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 RIGID BODY FIT CoLoRes, SITUS
The map was fitted using a model generated from the Nit structure (1ems in PDB). Magification was refined by dtermining the CoLoRes correlation coefficient as a function of scale factor. Locations of the 14 subunits were verified by vizualization with O. Fitted co-ordinates are available on request.Initially no symmetry was assumed but a two fold axis became apparent after 47 cycles. The orientation of the twofold axis was determined and the model was rotated so that the twofold axis was parallel to the x-axis. In 17 subsequent cycles of refinement the twofold symmetry was imposed by rotating about the x-axis and adding the structure thus formed to the original.