HPUB OBS RCSB RCSB RCSB 2013-01-16 2013-07-03 2014-03-19 2014-03-19 Electron cryo-microscopy of mutant p97R155P Mountassif D Fabre L Zaid Y Halawani D Bostina M Rouiller I p97, mutant, hIBMPFTD and Cryo-EM. Mountassif D Fabre L Zaid Y Halawani D Bostina M Rouiller I Impact of the mutation R155P implicated in hIBMPFTD diseases on the conformational change of D2-domain of the AAA ATPase p97/VCP To Be Published EMD-5574 FULLOVERLAP p97 R155P p97 R155P
The sample was very pure and homogenous.
homohexamer 1 0.6 0.6 Gel filtration and Polyacrylamide gel electrophoresis Transitional endoplasmic reticulum ATPase Mus musculus mouse endoplasmic reticulum cytoplasm 0.6 0.6 6 hexamer true Escherichia coli BL21(DE3) pTrcHis2C Q01853 singleParticle particle 0.2 7.5
20 mM Hepes, 150 mM NaCl, 5 mM MgCl2, 2.5 mM DTT
400 mesh copper grid
 ETHANE 100 120 FEI VITROBOT MARK IV
Vitrification carried out in ethane in liquid nitrogen atmosphere.
Blot 2 seconds, blot force 1 FEI TECNAI F20 FLOOD BEAM BRIGHT FIELD FIELD EMISSION GUN 200 1.0 3.5 50000.0 GATAN LIQUID NITROGEN 78 108 103 Objective lens astigmatism was corrected at 50,000 times magnification FEI 2012-02-14 GENERIC CCD 76 15 liquid nitrogen
Particles in multiple orientations were selected from cryo-micrographs using Signature and analyzed by XMipp software
ctffind3
 14.0 FSC 0.5 CUT-OFF Xmipp 15800 64 emd_5575.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 128 128 128 -64 -64 -64 128 128 128 294.4 294.4 294.4 90.0 90.0 90.0 X Y Z -0.03692038 0.05106695 0.00081872 0.0053219 2.3 2.3 2.3 0.029 EMDB Reconstruction of the mutant p97R155P
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 3CF1 C FLEXIBLE FIT Chimera, MDFF
The domains were symmetrically fitted into the map using the Molecular Dynamics Flexible Fitting software.
REAL