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Rapid Assembly of Alzheimer-like Paired Helical Filaments from Microtubule-Associated Protein Tau Monitored by Fluorescence in Solution
Abstract
Alzheimer's disease is characterized by the progressive deposition of two types of fibers in the affected brains, the amyloid fibers (consisting of the Abeta peptide, generating the amyloid plaques) and paired helical filaments (PHFs, made up of tau protein, forming the neurofibrillary tangles). While the principles of amyloid aggregation are known in some detail, the investigation of PHF assembly has been hampered by the low efficiency of tau aggregation, the requirement of high protein concentrations, and the lack of suitable detection methods. Here we report a quantitative assay system that permits monitoring of the assembly of PHFs in real time by the fluorescence of dyes such as thioflavine S or T. Using this assay, we evaluated parameters that influence the efficiency of filament formation. Disulfide-linked dimers of tau constructs representing the repeat domain assemble into PHFs most efficiently, but other tau isoforms or constructs form bona fide PHFs as well. The rate of assembly is greatly enhanced by polyanions such as RNA, heparin, and notably polyglutamate which resembles the acidic tail of tubulin. The assembly is optimal at pH approximately 6 and low ionic strengths (<50 mM) and increases steeply with temperatures above 30 degreesC, indicating that it is an entropy-driven process.
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“widely published aggregation data, which consistently reported more efficient fibrillization of K19/3R-Tau when compared to K18/4R-Tau23, 46, 48”
From Liquid–liquid phase separation of the microtubule-binding repeats of the Alzheimer-related protein Tau (2017) · cites this paper as [48]
In the citing paper
“which is based on the binding of ThT to cross-β-structure.”
From Liquid–liquid phase separation of the microtubule-binding repeats of the Alzheimer-related protein Tau (2017) · cites this paper as [48]
Citations
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Cited by
von Bergen, M., Friedhoff, P., Biernat, J., Heberle, J., Mandelkow, E.-M., & Mandelkow, E. (2000) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.97.10.5129 · Cited by 1,021
Self-citationAmbadipudi, S., Biernat, J., Riedel, D., Mandelkow, E., & Zweckstetter, M. (2017) · Nature Communications · doi:10.1038/s41467-017-00480-0 · Cited by 782
Self-citationCell-to-cell transmission of pathogenic proteins in neurodegenerative diseases
Guo, J. L., & Lee, V. M. Y. (2014) · Nature Medicine · doi:10.1038/nm.3457 · Cited by 641
Increasing O-GlcNAc slows neurodegeneration and stabilizes tau against aggregation
Yuzwa, S. A., Shan, X., Macauley, M. S., Clark, T., Skorobogatko, Y., Vosseller, K., & Vocadlo, D. J. (2012) · Nature Chemical Biology · doi:10.1038/nchembio.797 · Cited by 611
Proteasome inhibition by paired helical filament‐tau in brains of patients with Alzheimer's disease
Keck, S., Nitsch, R., Grune, T., & Ullrich, O. (2003) · Journal of Neurochemistry · doi:10.1046/j.1471-4159.2003.01642.x · Cited by 483
Metadata
- Authors
- Friedhoff, P., Schneider, A., Mandelkow, E.-M., & Mandelkow, E.
- Affiliations
- Max-Planck-Unit for Structural Molecular Biology, Notkestrasse 85, D-22607 Hamburg, Germany
- Venue
- Biochemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 1998-06-24
- Volume
- 37
- Issue
- 28
- Pages
- 10223-10230
- ISSN
- 0006-2960
- 1520-4995
- Cited by
- 373
- References
- 1
- Language
- en
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