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

N-terminal acetylation is one of the most common co- and post-translational modifications of the eukaryotic proteome and regulates numerous aspects of cellular physiology, such as protein folding, localization and turnover. In particular alpha-synuclein, whose dyshomeostasis has been tied to the pathogenesis of several neurodegenerative disorders, is completely N-alpha-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 P-BAD (L-arabinoseinducible) promoter. We show its functionality and the ability to completely N-alpha-acetylate our model substrate alpha-synuclein both upon induction of the construct with L-arabinose and also by only relying on the constitutive expression of the NatB genes.