Expression and purification of tau protein and its frontotemporal dementia variants using a cleavable histidine tag.
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.
School of Life Sciences, University of Warwick, Coventry CV4 7AL, UK; Midlands Integrative Biosciences Training Partnership, University of Warwick, Coventry CV4 7AL, UK. Electronic address: T.K.Karikari@warwick.ac.uk.
Thomas K
TK
Karikari
School of Life Sciences, University of Warwick, Coventry CV4 7AL, UK.
Alexandra
A
Turner
School of Life Sciences, University of Warwick, Coventry CV4 7AL, UK.
Robert
R
Stass
School of Life Sciences, University of Warwick, Coventry CV4 7AL, UK.
Leonie C Y
LC
Lee
School of Life Sciences, University of Warwick, Coventry CV4 7AL, UK.
Bethany
B
Wilson
Aston Research Center for Healthy Ageing, School of Life and Health Sciences, Aston University, Birmingham B4 7ET, UK.
David A
DA
Nagel
Aston Research Center for Healthy Ageing, School of Life and Health Sciences, Aston University, Birmingham B4 7ET, UK.
Eric J
EJ
Hill
School of Life Sciences, University of Warwick, Coventry CV4 7AL, UK.
Kevin G
KG
Moffat
D002846:Chromatography, Affinity / Q000379:methods; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D057180:Frontotemporal Dementia; D006639:Histidine / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D016875:tau Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification
D011993:Recombinant Fusion Proteins; D016875:tau Proteins; C033223:polyhistidine; D006639:Histidine
Alzheimer's disease; Expression; FTPD-17; Microtubule-associated protein tau; Purification; Tau; Tauopathies
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