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Recombinant production and purification of the human protein Tau
Protein Engineering, Design and Selection ( IF 2.4 ) Pub Date : 2019-07-02 , DOI: 10.1093/protein/gzz010
Luca Ferrari 1, 2 , Stefan G D Rüdiger 1, 2
Affiliation  

Tau protein is a microtubule-stabilising protein whose aggregation is linked to Alzheimer’s Disease and other forms of dementia. Tau biology is at the heart of cytoskeletal dynamics and neurodegenerative mechanisms, making it a crucial protein to study. Tau purification, however, is challenging as Tau is disordered, which makes it difficult to produce in recombinant system and is degradation-prone. It is thus challenging to obtain pure and stable preparations of Tau. Here, we present a fast and robust protocol to purify Tau recombinantly in Escherichia coli. Our protocol allows purifying Tau either tag-less or FLAG-tagged at its N-terminus, and Tau fragments of interest. By exploiting a cleavable affinity tag and two anion exchange columns, we obtained Tau preparations of high purity, stable and suitable for in vitro studies, including aggregation experiments that resemble neurodegenerative processes.

中文翻译:

人蛋白Tau的重组生产和纯化

Tau蛋白是一种微管稳定蛋白,其聚集与阿尔茨海默氏病和其他形式的痴呆症有关。Tau生物学是细胞骨架动力学和神经退行性机制的核心,使其成为研究的关键蛋白质。然而,由于Tau无序,所以Tau纯化具有挑战性,这使其难以在重组系统中生产并且易于降解。因此,获得纯和稳定的Tau制剂是具有挑战性的。在这里,我们提出了一种快速,可靠的方案,以在大肠杆菌中重组纯化Tau。我们的协议允许在其N末端纯化无标签或带有FLAG标签的Tau,以及感兴趣的Tau片段。通过利用可裂解的亲和标签和两个阴离子交换柱,我们获得了高纯度,稳定且适合体外研究的Tau制剂,
更新日期:2019-07-23
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