An efficient method for the purification of proteins from four distinct toxin-antitoxin modules.

TitleAn efficient method for the purification of proteins from four distinct toxin-antitoxin modules.
Publication TypeJournal Article
Year of Publication2015
AuthorsSterckx, Y. G. - J., S. De Gieter, V. Zorzini, S. Hadži, S. Haesaerts, R. Loris, and A. Garcia-Pino
JournalProtein Expr Purif
Volume108
Pagination30-40
Date Published2015 Apr
ISSN1096-0279
KeywordsArchaeal Proteins, Bacterial Proteins, Multiprotein Complexes
Abstract

Toxin-antitoxin (TA) modules are stress response elements that are ubiquitous in the genomes of bacteria and archaea. Production and subsequent purification of individual TA proteins is anything but straightforward as over-expression of the toxin gene is lethal to bacterial and eukaryotic cells and over-production of the antitoxin leads to its proteolytic degradation because of its inherently unstructured nature. Here we describe an effective production and purification strategy centered on an on-column denaturant-induced dissociation of the toxin-antitoxin complex. The success of the method is demonstrated by its application on four different TA families, encoding proteins with distinct activities and folds. A series of biophysical and in vitro activity tests show that the purified proteins are of high quality and suitable for structural studies.

DOI10.1016/j.pep.2015.01.001
Alternate JournalProtein Expr. Purif.
PubMed ID25582764
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