2023-06-14RELRCSBRCSBRCSB2020-03-022020-04-012021-01-132023-06-14Amyloid-beta(1-40) fibril derived from Alzheimer's disease cortical tissueGhosh UThurber KRTycko Ramyloid-beta, Alzheimer's disease, PROTEIN FIBRILGhosh UThurber KRYau WMTycko RMolecular structure of a prevalent amyloid-beta fibril polymorph from Alzheimer's disease brain tissue.Proc.Natl.Acad.Sci.USAUS118202133431654doi:10.1073/pnas.20230891181091-64900040PNASA6EMD-21501associated EM volumeAmyloid-beta(1-40) fibril derived from Alzheimer's disease cortical tissue6w0oFULLOVERLAPamyloid-beta(1-40) fibrils derived from human AD brainamyloid-beta(1-40) fibrils derived from human AD brain01Fibrils produced by seeded growth using amyloid-beta in brain extract as the source of seeds. CryoEM and solid state NMR measurements were performed on second-generation seeded fibrils.Homo sapiens29Amyloid-beta precursor proteinHomo sapiens0.0043358519999999996LEVODAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVhelicalfilament0.457.410.0Na2HPO4/NaH2PO4Phosphate buffer10 mM phosphate buffer with 0.01% NaN3 to avoid microbial contamination. Buffers were filtered to avoid contamination.Quantifoil R1.2/1.3GOLD300HOLEYGLOW DISCHARGEOTHER0.036000000000000004The grids were checked in microscope prior to use.ETHANE90293LEICA PLUNGERThe grids were preblotted for 10 seconds and blotted for 6 seconds before plunging.. Protein exists in solution as amyloid fibrils of varying lengths.TFS KRIOSFLOOD BEAMBRIGHT FIELDFIELD EMISSION GUN300100.02.7-0.5-3.0130000.0FEI TITAN KRIOS AUTOGRID HOLDERNITROGEN6.0GIF Quantum LS20Preliminary grid screening was done manually in FEI T12.GATAN K2 QUANTUM (4k x 4k)SUPER-RESOLUTION1-501133710.073.51The selected images were corrected for gain. The gain reference was not rotated or flipped.12.45179.66C1FOURIER SPACE2.77FSC 0.143 CUT-OFFRELIONcustommodified from 3.0-betaA modified version of RELION 3.0-beta was used for the reconstruction. The modified version allowed local averaging of the rotation angle. Final reconstruction used 2-fold screw symmetry.1477519790RELION3.0manual pickingThe particles were picked using Relion manual pick, with a 20A low pass filter and particle diameter of 30A.The initial model was taken from a lower-resolution cryoEM reconstruction of amyloid-beta(1-40) fibrils prepared without brain seeding, low-pass filtered at 10 Angstrom resolution.NOT APPLICABLEAB INITIO MODELXplor-NIH was used to combine EM density with phi/psi restraints from NMR chemical shifts (from Talos-N Version 4.21 Rev 2016.343.11.31).REALemd_21501_msk_1.map