Preparation of stable tau oligomers for cellular and biochemical studies

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.

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.

Publication DOI: https://doi.org/10.1016/j.ab.2018.10.013
Divisions: Life & Health Sciences > Biosciences
Life & Health Sciences
Additional Information: © 2018 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/BY/4.0/) 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
Full Text Link:
Related URLs: https://linking ... 003269718305244 (Publisher URL)
http://www.scop ... tnerID=8YFLogxK (Scopus URL)
Published Online Date: 2019-02-01
Published Date: 2019-02-01
Authors: Karikari, Thomas K.
Nagel, David A.
Grainger, Alastair
Clarke-bland, Charlotte
Hill, Eric J. ( 0000-0002-9419-1500)
Moffat, Kevin G.

Download

[img]

Version: Published Version

License: Creative Commons Attribution

| Preview

Export / Share Citation


Statistics

Additional statistics for this record