2024-05-22 REL PDBe PDBe PDBe 2019-11-27 2020-02-05 2020-02-05 2024-05-22 Medical Research Council (United Kingdom) MC_UP_A025_1013 United Kingdom Medical Research Council (United Kingdom) MC_U105184291 United Kingdom Zhang W Murzin AG Falcon B Shi Y Goedert M Scheres SHW tau protein, filament, cross-beta structure, PROTEIN FIBRIL Zhang W Tarutani A Newell KL Murzin AG Matsubara T Falcon B Vidal R Garringer HJ Shi Y Ikeuchi T Murayama S Ghetti B Hasegawa M Goedert M Scheres SHW Nature UK 580 283 287 2020 32050258 doi:10.1038/s41586-020-2043-0 1476-4687 0006 NATUAS EMD-10512 other EM volume

The map of different filaments from one dataset

6tjx FULLOVERLAP Tau filaments extracted from the Frontal cortex of a patient with corticobasal degeneration. Tau filaments extracted from the Frontal cortex of a patient with corticobasal degeneration. 0 1

Corticobasal degeneration (CBD) is characterised by abundant filamentous tau inclusions that are made of isoforms with four microtubule-binding repeats (4R).

Homo sapiens 0.46 Microtubule-associated protein tau Homo sapiens 0.045919870999999994 6 LEVO MAEPRQEFEVMEDHAGTYGLGDRKDQGGYTMHQDQEGDTDAGLKESPLQTPTEDGSEEPGSETSDAKSTPTAEDVTAPLV DEGAPGKQAAAQPHTEIPEGTTAEEAGIGDTPSLEDEAAGHVTQARMVSKSKDGTGSDDKKAKGADGKTKIATPRGAAPP GQKGQANATRIPAKTPPAPKTPPSSGEPPKSGDRSGYSSPGSPGTPGSRSRTPSLPTPPTREPKKVAVVRTPPKSPSSAK SRLQTAPVPMPDLKNVKSKIGSTENLKHQPGGGKVQIINKKLDLSNVQSKCGSKDNIKHVPGGGSVQIVYKPVDLSKVTS KCGSLGNIHHKPGGGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGGNKKIETHKLTFRENAKAKTDHGAEIVYKSPVVS GDTSPRHLSNVSSTGSIDMVDSPQLATLADEVSASLAKQGL P10636 helical filament 2.0 7.4 0.02 Tris tris(hydroxymethyl)aminomethane 0.1 NaCl sodium chloride

20 mM Tris, pH 7.4, 100mM NaCl

ETHANE 100 277 FEI VITROBOT MARK IV

Blot force: -12 ; Blot time: 4s.

The tau filaments were extracted from the Frontal cortex of a patient with corticobasal degeneration by using sarkosyl and ultracentrifuge.

FEI TITAN KRIOS FLOOD BEAM BRIGHT FIELD FIELD EMISSION GUN 300 70.0 2.7 1.7 2.8000000000000003 FEI TITAN KRIOS AUTOGRID HOLDER NITROGEN GIF Quantum LS 20 GATAN K2 SUMMIT (4k x 4k) COUNTING 3838 3710 1-40 1 2567 10.0 1.346

Images were collected in movie-mode at 40 frames every 10 seconds

1

Movie frames were gain-corrected, aligned, dose weighted and then summed into a single micrograph using MOTIONCOR2 (Zheng et al., 2017)

1 4.786 -0.61 C2 3.0 FSC 0.143 CUT-OFF RELION 3.0 20752 129812 RELION 3.0

Manually picked

An initial 3D reference was reconstructed from the 2D class averages de novo.

NOT APPLICABLE RELION 3.0 AB INITIO MODEL

A stack of three consecutive monomers was refined to preserve nearest-neighbour interactions for the middle chain. Because most residues adopted cross strand conformation, hydrogen-bond restraints were imposed to preserve a parallel, in-register hydrogen bonding pattern in earlier stages of Fourier-space refinements. Local symmetry restraints were imposed to keep all beta strand rungs identical. 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.

Fourier shell correlation RECIPROCAL 25.75 emd_10514_msk_1.map emd_10514_half_map_1.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 330 330 330 0 0 0 330 330 330 379.5 379.5 379.5 90.0 90.0 90.0 X Y Z -0.026520643 0.060516972 0.0005823076 0.0031659503 1.15 1.15 1.15 AUTHOR ::::EMDATABANK.org::::EMD-10514:::: Half map1 emd_10514_half_map_2.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 330 330 330 0 0 0 330 330 330 379.5 379.5 379.5 90.0 90.0 90.0 X Y Z -0.02296103 0.058970653 0.0005827394 0.0031025005 1.15 1.15 1.15 AUTHOR ::::EMDATABANK.org::::EMD-10514:::: Half map2