Use of the myosin motor domain as large-affinity tag for the expression and purification of proteins in Dictyostelium discoideum

International Journal of Biological Macromolecules
Martin Kollmar

Abstract

The cellular slime mold Dictyostelium discoideum is increasingly be used for the overexpression of proteins. Dictyostelium is amenable to classical and molecular genetic approaches and can easily be grown in large quantities. It contains a variety of chaperones and folding enzymes, and is able to perform all kinds of post-translational protein modifications. Here, new expression vectors are presented that have been designed for the production of proteins in large quantities for biochemical and structural studies. The expression cassettes of the most successful vectors are based on a tandem affinity purification tag consisting of an octahistidine tag followed by the myosin motor domain tag. The myosin motor domain not only strongly enhances the production of fused proteins but is also used for a fast affinity purification step through its ATP-dependent binding to actin. The applicability of the new system has been demonstrated for the expression and purification of subunits of the dynein-dynactin motor protein complex from different species.

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Citations

Aug 31, 2011·Journal of Molecular Biology·Rebecca J MoenMargaret A Titus
Oct 3, 2014·PloS One·Mónica González-MagaldiFrancisco Sobrino
Aug 21, 2008·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Ranjana AryaKulvinder Singh Saini
Mar 6, 2008·Langmuir : the ACS Journal of Surfaces and Colloids·Rafael Tadmor, Ken G Pepper
Jul 25, 2021·International Journal of Molecular Sciences·Laura M Nederveen-SchippersVictor Skakun

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