Open AccessPick-ya actin: a method to purify actin isoforms with bespoke key post-translational modifications
Author(s) -
Tomoyuki Hatano,
Lavanya Sivashanmugam,
Andrejus Suchenko,
Hamdi Hussain,
Mohan K. Balasubramanian
Publication year - 2020
Publication title -
journal of cell science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.384
H-Index - 278
eISSN - 1477-9137
pISSN - 0021-9533
DOI - 10.1242/jcs.241406
Subject(s) - biology , actin , eukaryote , gene isoform , acetylation , microbiology and biotechnology , cytoskeleton , biochemistry , cofilin , actin cytoskeleton , actin binding protein , profilin , actin remodeling , cell , gene , genome
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.