9BBL

THF filament generated from 4E-Tau(297-407) under neutral Mg2+ condition

Summary for 9BBL

• Entry DOI: 10.2210/pdb9bbl/pdb
• EMDB information: 43824 43826 44421
• Descriptor: Isoform Tau-F of Microtubule-associated protein tau (1 entity in total)
• Functional Keywords: tau, amyloid fibrils, in vitro, phf, protein fibril
• Biological source: Homo sapiens (human)
• Total number of polymer chains: 9
• Total formula weight: 414665.89

Authors

Duan, P.,El Mammeri, N. (deposition date: 2024-04-06, release date: 2024-05-08, Last modification date: 2024-06-05)

Primary citation

Duan, P.,El Mammeri, N.,Hong, M.
Milligram-scale assembly and NMR fingerprint of tau fibrils adopting the Alzheimer's disease fold.
J.Biol.Chem., 300:107326-107326, 2024

PubMed Abstract: In the Alzheimer's disease (AD) brain, the microtubule-associated protein tau aggregates into paired helical filaments in which each protofilament has a C-shaped conformation. In vitro assembly of tau fibrils adopting this fold is highly valuable for both fundamental and applied studies of AD without requiring patient-brain extracted fibrils. To date, reported methods for forming AD-fold tau fibrils have been irreproducible and sensitive to subtle variations in fibrillization conditions. Here, we describe a route to reproducibly assemble tau fibrils adopting the AD fold on the multi-milligram scale. We investigated the fibrillization conditions of two constructs and found that a tau (297-407) construct that contains four AD phospho-mimetic glutamate mutations robustly formed the C-shaped conformation. 2D and 3D correlation solid-state NMR spectra show a single predominant set of chemical shifts, indicating a single molecular conformation. Negative-stain electron microscopy and cryo-EM data confirm that the protofilament formed by 4E-tau (297-407) adopts the C-shaped conformation, which associates into paired, triple, and quadruple helical filaments. In comparison, NMR spectra indicate that a previously reported construct, tau (297-391), forms a mixture of a four-layered dimer structure and the C-shaped structure, whose populations are sensitive to the environmental conditions. The determination of the NMR chemical shifts of the AD-fold tau opens the possibility for future studies of tau fibril conformations and ligand binding by NMR. The quantitative assembly of tau fibrils adopting the AD fold should facilitate the development of diagnostic and therapeutic compounds that target AD tau.

PubMed: 38679331
DOI: 10.1016/j.jbc.2024.107326

Experimental method

ELECTRON MICROSCOPY (2.5 Å)