Expression and purification of tau protein and its frontotemporal dementia variants using a cleavable histidine tag

Abstract

Recombinant tau protein is widely used to study the biochemical, cellular and pathological aspects of tauopathies, including Alzheimer's disease and frontotemporal dementia with Parkinsonism linked to chromosome 17 (FTPD-17). Pure tau in high yield is a requirement for in vitro evaluation of the protein's physiological and toxic functions. However, the preparation of recombinant tau is complicated by the protein's propensity to aggregate and form truncation products, necessitating the use of multiple, time-consuming purification methods. In this study, we investigated parameters that influence the expression of wild type and FTPD-17 pathogenic tau, in an attempt to identify ways to maximise expression yield. Here, we report on the influence of the choice of host strain, induction temperature, duration of induction, and media supplementation with glucose on tau expression in Escherichia coli. We also describe a straightforward process to purify the expressed tau proteins using immobilised metal affinity chromatography, with favourable yields over previous reports. An advantage of the described method is that it enables high yield production of functional oligomeric and monomeric tau, both of which can be used to study the biochemical, physiological and toxic properties of the protein.

Publication DOI: https://doi.org/10.1016/j.pep.2016.09.009
Divisions: Life & Health Sciences > Biosciences
Life & Health Sciences > Chronic and Communicable Conditions
Life & Health Sciences
Life & Health Sciences > Clinical and Systems Neuroscience
Life & Health Sciences > Cell & Tissue Biomedical Research
Additional Information: © 2016 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 (NC3Rs)’s CRACKIT: Untangle (NC/C013101/1); University of Warwick Chancellor's Scholarship; and BBSRC (BB/J014532/1). Supplementarydatarelatedtothisarticlecanbefoundathttp://dx.doi.org/10.1016/j.pep.2016.09.009.
Uncontrolled Keywords: Alzheimer's disease,expression,FTPD-17,microtubule-associated protein tau,purification,tau,tauopathies,Biotechnology
Full Text Link:
Related URLs: http://www.scop ... tnerID=8YFLogxK (Scopus URL)
PURE Output Type: Article
Published Date: 2017-02
Published Online Date: 2016-09-20
Accepted Date: 2016-09-19
Submitted Date: 2016-08-24
Authors: Karikari, Thomas K.
Turner, Alexandra
Stass, Robert
Lee, Leonie C.Y.
Wilson, Bethany
Nagel, David A.
Hill, Eric J. ( 0000-0002-9419-1500)
Moffat, Kevin G.

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