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

Autor:	Remy Loris, Valentina Zorzini, Steven De Gieter, Abel Garcia-Pino, San Hadži, Yann G.-J. Sterckx, Sarah Haesaerts
Rok vydání:	2015
Předmět:	biology Toxin Archaeal Proteins biology.organism_classification medicine.disease_cause Genome In vitro Microbiology Bacterial Proteins Biochemistry Structural biology Multiprotein Complexes Protein purification medicine Antitoxin Gene Bacteria Biotechnology
Zdroj:	Protein Expression and Purification. 108:30-40
ISSN:	1046-5928
DOI:	10.1016/j.pep.2015.01.001
Popis:	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.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::914225272b4ddccf354fab71295bb7fe https://doi.org/10.1016/j.pep.2015.01.001 Zobrazit plný text záznamu Full Text from ScienceDirect