Karikari, Thomas K., Turner, Alexandra, Stass, Robert, Lee, Leonie C.Y., Wilson, Bethany, Nagel, David A., Hill, Eric J. and Moffat, Kevin G. (2016) Expression and purification of tau protein and its frontotemporal dementia variants using a cleavable histidine tag. Protein Expression and Purification, 130 . pp. 44-54. doi:10.1016/j.pep.2016.09.009 ISSN 1046-5928.

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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.

Item Type:	Journal Article
Subjects:	R Medicine > RC Internal medicine T Technology > TP Chemical technology
Divisions:	Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- )
Library of Congress Subject Headings (LCSH):	Recombinant proteins, Alzheimer's disease, Dementia, Parkinson's disease
Journal or Publication Title:	Protein Expression and Purification
Publisher:	Academic Press
ISSN:	1046-5928
Official Date:	February 2016
Dates:	Date Event February 2016 Published 19 September 2016 Accepted 20 September 2016 Available 24 August 2016 Submitted
Volume:	130
Page Range:	pp. 44-54
DOI:	10.1016/j.pep.2016.09.009
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Restricted or Subscription Access
Date of first compliant deposit:	10 October 2016
Date of first compliant Open Access:	11 October 2016
Funder:	National Centre for the Replacement, Refinement, and Reduction of Animals in Research (Great Britain) (NC3R), University of Warwick Chancellor's Scholarship, Biotechnology and Biological Sciences Research Council (Great Britain) (BBSRC)
Grant number:	NC/C013101/1 (NC3R), B/J014532/1 (BBSRC)
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