Structural Characterization of Amyloid-β Fibrils

Full details of sample preparations, measurements, data analyses, and simulations are given in SI Appendix. Briefly, second-generation Aβ42 fibrils were grown from synthetic or recombinant peptide, using sonicated first-generation brain-seeded fibrils described by Qiang et al. (5 (link)) as seeds. TEM and AFM images were obtained as previously described (5 (link), 24 (link)). Cryo-EM grids were screened with a Thermo Fisher Glacios microscope. Final images were obtained with a Thermo Fisher Titan Krios microscope. Cryo-EM images were processed and density maps were reconstructed with RELION software (18 (link), 24 (link)). ssNMR data were obtained at 14.1 T and 17.5 T, using Tecmag Redstone spectrometers, magic-angle spinning probes obtained from the research group of Drs. Ago Samoson and from Black Fox, LLC, and standard pulse sequence methods. MD simulations were performed with NAMD software and analyzed with VMD software (38 (link), 39 (link)).

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Lee M., Yau W.M., Louis J.M, & Tycko R. (2023). Structures of brain-derived 42-residue amyloid-β fibril polymorphs with unusual molecular conformations and intermolecular interactions. Proceedings of the National Academy of Sciences of the United States of America, 120(11), e2218831120.

Publication 2023

Glacios microscope Titan Krios microscope

Brain Maps Microscope Peptide Pulse Seeds

Corresponding Organization :

Other organizations : National Institute of Diabetes and Digestive and Kidney Diseases

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Variable analysis

independent variables

Concentration of synthetic or recombinant Aβ42 peptide
Sonicated first-generation brain-seeded fibrils as seeds for growing second-generation Aβ42 fibrils

dependent variables

Structural characteristics of Aβ42 fibrils (as observed by TEM, AFM, and cryo-EM)
SsNMR data of Aβ42 fibrils
Results of MD simulations of Aβ42 fibrils

control variables

Experimental methods and protocols for TEM, AFM, cryo-EM, ssNMR, and MD simulations (as described in the SI Appendix)

controls

Sonicated first-generation brain-seeded fibrils described by Qiang et al. (2017) as positive control for growing second-generation Aβ42 fibrils

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