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Preparation of stable tau oligomers for cellular and biochemical studies

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Karikari, Thomas K., Nagel, David A., Grainger, Alastair, Clarke-Bland, Charlotte, Hill, Eric J. and Moffat, Kevin G. (2019) Preparation of stable tau oligomers for cellular and biochemical studies. Analytical Biochemistry, 566 . pp. 67-74. doi:10.1016/j.ab.2018.10.013 ISSN 0003-2697.

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Official URL: http://doi.org/10.1016/j.ab.2018.10.013

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Abstract

Increasing evidence suggests that small oligomers are the principal neurotoxic species of tau in Alzheimer's disease and other tauopathies. However, mechanisms of tau oligomer-mediated neurodegeneration are poorly understood. The transience of oligomers due to aggregation can compromise the stability of oligomers prepared in vitro. Consequently, we sought to develop an efficient method which maintains the stability and globular conformation of preformed oligomers. This study demonstrates that labeling a single-cysteine form of the pro-aggregant tau four-repeat region (K18) with either Alexa Fluor 488-C5-maleimide or N-ethylmaleimide in reducing conditions stabilizes oligomers by impeding their further aggregation. Furthermore, the use of this approach to study the propagation of labeled extracellular tau K18 oligomers into human neuroblastoma cells and human stem cell-derived neurons is described. This method is potentially applicable for preparing stabilized oligomers of tau for diagnostic and biomarker tests, as well as for in vitro structure-activity relationship assays.

Item Type: Journal Article
Subjects: R Medicine > RC Internal medicine > RC0321 Neuroscience. Biological psychiatry. Neuropsychiatry
Divisions: Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- )
Library of Congress Subject Headings (LCSH): Alzheimer's disease -- Research, Oligomers, Neurotoxic agents
Journal or Publication Title: Analytical Biochemistry
Publisher: Academic Press
ISSN: 0003-2697
Official Date: 1 February 2019
Dates:
DateEvent
1 February 2019Published
11 October 2018Available
8 October 2018Accepted
Volume: 566
Page Range: pp. 67-74
DOI: 10.1016/j.ab.2018.10.013
Status: Peer Reviewed
Publication Status: Published
Reuse Statement (publisher, data, author rights): © 2018, Elsevier. Licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/.
Access rights to Published version: Open Access (Creative Commons)
Date of first compliant deposit: 16 October 2018
Date of first compliant Open Access: 11 October 2019
RIOXX Funder/Project Grant:
Project/Grant IDRIOXX Funder NameFunder ID
NC/C013101/1National Centre for the Replacement, Refinement and Reduction of Animals in Researchhttp://dx.doi.org/10.13039/501100000849
BB/J014532/1[BBSRC] Biotechnology and Biological Sciences Research Councilhttp://dx.doi.org/10.13039/501100000268
UNSPECIFIEDUniversity of Warwick UNSPECIFIED
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