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

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• 作者：Thomas K. Karikari^a ; ^b ; ^{T.K.Karikari@warwick.ac.uk} ; Alexandra Turner^a ; Robert Stass^a ; Leonie C.Y. Lee^a ; Bethany Wilson^a ; David A. Nagel^c ; Eric J. Hill^c ; Kevin G. Moffat^a
• 关键词：Tau ; Tauopathies ; Microtubule-associated protein tau ; Alzheimer's disease ; FTPD-17 ; Expression ; Purification
• 刊名：Protein Expression and Purification
• 出版年：2017
• 出版时间：February 2017
• 年：2017
• 卷：130
• 期：Complete
• 页码：44-54
• 全文大小：1575 K
• 卷排序：130

文摘

Factors influencing the expression of wild type and FTPD-17 pathogenic tau were investigated in an attempt to maximise yield. Soluble monomeric tau expression level was highest at 37 °C compared to 20 °C and 25 °C. Media supplementation with 0.2% glucose did not significantly influence monomeric tau expression levels. Circular dichroism confirmed that the purified tau proteins were mostly unfolded, with negative peaks around 200 nm. The circular dichroism peaks shifted towards 220 nm following the preparation of Alzheimer-like filaments, suggesting secondary structure re-orientation towards β-sheets. The tau proteins adopted classical fibrillisation similar to filaments isolated from the brains of Alzheimer's disease patients.