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bioRxiv · 10.1101/331355

pTSara-NatB, an improved N-terminal acetylation system for recombinant protein expression in E. coli

Rovere, M.·Powers, A. E.·Patel, D. S.·Bartels, T.
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Abstract

N-terminal acetylation is one of the most common post-translational modifications of the eukaryotic proteome and regulates numerous aspects of cellular physiology, such as protein folding, localization and turnover. In particular -synuclein, whose dyshomeostasis has been tied to the pathogenesis of several neurodegenerative disorders, is completely N-acetylated in nervous tissue. In this work, building on previous reports, we develop and characterize a bacterial N-terminal acetylation system based on the expression of the yeast N-terminal acetyltransferase B (NatB) complex under the control of the PBAD (L-arabinose-inducible) promoter. We show its functionality and the ability to completely N-acetylate our model substrate -synuclein both upon induction of the construct with L-arabinose and also by only relying on the constitutive expression of the NatB genes.

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Rovere, M., Powers, A. E., Patel, D. S., Bartels, T.. 2018-05-25. pTSara-NatB, an improved N-terminal acetylation system for recombinant protein expression in E. coli. https://doi.org/10.1101/331355

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