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bioRxiv · 10.1101/833152

Pick-ya actin: A method to purify actin isoforms with bespoke key post-translational modifications

Hatano, T.·Sivashanmugam, L.·Suchenko, A.·Hussain, H.·Balasubramanian, M.
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Abstract

Actin is one of the most abundant eukaryotic cytoskeletal polymer-forming proteins, which in the filamentous form regulates a number of physiological processes, ranging from cell division and migration to development and tissue function. Actins are differentially post-translationally modified (PTMs) in different organisms, which include Met, Ala, Asp, and Glu N-acetylation, N-arginylation, and the 73th His residue (His-73) methylation, with different organisms displaying a distinct signature of PTMs. Currently methods are not available to produce actin isoforms with organism specific PTM profile. Here we report Pick-ya actin, a method to express actin isoforms from any eukaryote with its own key characteristic PTM pattern. We achieve this using a synthetic biology strategy in a yeast strain that expresses 1. actin isoforms with the desired N-end via ubiquitin fusion and 2. mammalian enzymes that promote acetylation and methylation. Pick-ya actin should greatly facilitate biochemical, structural, and physiological studies of the actin cytoskeleton and its PTMs.

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Hatano, T., Sivashanmugam, L., Suchenko, A., Hussain, H., Balasubramanian, M.. 2019-11-06. Pick-ya actin: A method to purify actin isoforms with bespoke key post-translational modifications. https://doi.org/10.1101/833152

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