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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
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 | ||||||||||||
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| 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: |
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| 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: |
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