2019-03-27 REL 2019-01-22 PDBe PDBe 2019-02-20 2019-02-20 Cryo-EM reconstruction of heparin-induced 2N4R tau snake filaments Zhang W, Falcon B, Murzin AG, Fan J, Crowther RA, Goedert M, Scheres SHW 6qjh Zhang W, Falcon B, Murzin AG, Fan J, Crowther RA, Goedert M, Scheres SH Heparin-induced tau filaments are polymorphic and differ from those in Alzheimer's and Pick's diseases. Elife 8 2019 30720432 doi:10.7554/eLife.43584 2050-084X emd_4563.map.gz Image stored as Reals 256 256 256 0.0 0.0 0.0 255.0 255.0 255.0 256 256 256 353.28 353.28 353.28 90.0 90.0 90.0 X Y Z -0.25210795 0.50170285 0.0005556408 0.0146968 1
1.38 1.38 1.38 0.0475 The helical reconstruction of heparin-induced 2N4R tau snake filaments 2 heparin-induced 2N4R tau snake filaments
Recombinant 2N4R tau protein was induced into filaments by adding heparin
protein heparin-induced 2N4R tau snake filaments 45.85
Recombinant 2N4R tau protein was induced into filaments by adding heparin
Homo sapiens false Escherichia coli BL21(DE3) pRK172 protein Isoform Tau-F of Microtubule-associated protein tau Homo sapiens false Escherichia coli BL21(DE3) ETHANE 100 277 FEI VITROBOT MARK IV
Blot force: -12 ; Blot time: 4s.
FIELD EMISSION GUN 48.0 BRIGHT FIELD 1700.0 1700.0 FLOOD BEAM FEI FALCON III (4k x 4k) 2.0 FEI POLARA 300 GATAN LIQUID NITROGEN 300 717 14.0
Images were collected in movie-mode at 30 frames per second
 Fourier shell correlation 41.259999999999998 RECIPROCAL
A stack of three consecutive monomers was refined to preserve nearest-neighbour interactions for the middle chain. Side-chain clashes were detected using MOLPROBITY, and corrected by iterative cycles of real-space refinement in COOT and Fourier-space refinement in REFMAC and PHENIX. For each refined structure, separate model refinements were performed against a single half-map, and the resulting model was compared to the other half-map to confirm the absence of overfitting.
 filament 2.0 7.4
20 mM Tris, pH 7.4, 100mM NaCl
 -1.26 4.7 C1 helical RELION Aligned, non-dose-weighted micrographs were used to estimate the contrast transfer function (CTF) using CTFFIND4.1 3.3 FSC 0.143 CUT-OFF
Movie frames were gain-corrected, aligned, dose weighted and then summed into a single micrograph using MOTIONCOR2 (Zheng et al., 2017)