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

Tomoyuki Hatano, Lavanya Sivashanmugam, Andrejus Suchenko, Hamdi Hussain, View ORCID ProfileMohan K. Balasubramanian
doi: https://doi.org/10.1101/833152
Tomoyuki Hatano
Centre for Mechanochemical Cell Biology and Division of Biomedical Sciences, Warwick Medical School, University of Warwick, Coventry CV4 7AL, United Kingdom
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  • For correspondence: T.Hatano@warwick.ac.uk M.K.Balasubramanian@warwick.ac.uk
Lavanya Sivashanmugam
Centre for Mechanochemical Cell Biology and Division of Biomedical Sciences, Warwick Medical School, University of Warwick, Coventry CV4 7AL, United Kingdom
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Andrejus Suchenko
Centre for Mechanochemical Cell Biology and Division of Biomedical Sciences, Warwick Medical School, University of Warwick, Coventry CV4 7AL, United Kingdom
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Hamdi Hussain
Centre for Mechanochemical Cell Biology and Division of Biomedical Sciences, Warwick Medical School, University of Warwick, Coventry CV4 7AL, United Kingdom
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Mohan K. Balasubramanian
Centre for Mechanochemical Cell Biology and Division of Biomedical Sciences, Warwick Medical School, University of Warwick, Coventry CV4 7AL, United Kingdom
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  • ORCID record for Mohan K. Balasubramanian
  • For correspondence: T.Hatano@warwick.ac.uk M.K.Balasubramanian@warwick.ac.uk
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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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Posted November 06, 2019.
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Pick-ya actin: A method to purify actin isoforms with bespoke key post-translational modifications
Tomoyuki Hatano, Lavanya Sivashanmugam, Andrejus Suchenko, Hamdi Hussain, Mohan K. Balasubramanian
bioRxiv 833152; doi: https://doi.org/10.1101/833152
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Pick-ya actin: A method to purify actin isoforms with bespoke key post-translational modifications
Tomoyuki Hatano, Lavanya Sivashanmugam, Andrejus Suchenko, Hamdi Hussain, Mohan K. Balasubramanian
bioRxiv 833152; doi: https://doi.org/10.1101/833152

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