Preparation of stable tau oligomers for cellular and biochemical studies


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.

Publication DOI:
Divisions: College of Health & Life Sciences > School of Biosciences
College of Health & Life Sciences > School of Biosciences > Cell & Tissue Biomedical Research
College of Health & Life Sciences
College of Health & Life Sciences > Chronic and Communicable Conditions
College of Health & Life Sciences > Clinical and Systems Neuroscience
Additional Information: © 2018 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license ( Funding: National Centre for the Replacement, Refinement and Reduction of Animals in Research (grant NC/C013101/1); Biotechnology and Biological Sciences Research Council grant number BB/J014532/1.
Uncontrolled Keywords: Alzheimer's disease,iPSC-derived neurons,Maleimide,Oligomer,Tau,Tauopathies,Biophysics,Biochemistry,Molecular Biology,Cell Biology
Publication ISSN: 1096-0309
Last Modified: 29 Apr 2024 07:27
Date Deposited: 18 Oct 2018 08:14
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Related URLs: https://linking ... 003269718305244 (Publisher URL)
http://www.scop ... tnerID=8YFLogxK (Scopus URL)
PURE Output Type: Article
Published Date: 2019-02-01
Published Online Date: 2018-10-11
Accepted Date: 2018-10-08
Authors: Karikari, Thomas K.
Nagel, David A. (ORCID Profile 0000-0002-9055-1775)
Grainger, Alastair
Clarke-bland, Charlotte (ORCID Profile 0000-0001-5776-2733)
Hill, Eric J. (ORCID Profile 0000-0002-9419-1500)
Moffat, Kevin G.



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License: Creative Commons Attribution

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License: Creative Commons Attribution Non-commercial No Derivatives

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