title: An efficient method for the purification of proteins from four distinct toxin-antitoxin modules.
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.
authors.affiliation: Structural Biology Brussels, Department of Biotechnology, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium; Structural Biology Research Center, VIB, Pleinlaan 2, B-1050 Brussels, Belgium.
authors.forename: Yann G-J
authors.initials: YG
authors.lastname: Sterckx
authors.affiliation: Structural Biology Brussels, Department of Biotechnology, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium; Structural Biology Research Center, VIB, Pleinlaan 2, B-1050 Brussels, Belgium.
authors.forename: Steven
authors.initials: S
authors.lastname: De Gieter
authors.affiliation: Structural Biology Brussels, Department of Biotechnology, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium; Structural Biology Research Center, VIB, Pleinlaan 2, B-1050 Brussels, Belgium.
authors.forename: Valentina
authors.initials: V
authors.lastname: Zorzini
authors.affiliation: Structural Biology Brussels, Department of Biotechnology, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium; Structural Biology Research Center, VIB, Pleinlaan 2, B-1050 Brussels, Belgium; Department of Physical Chemistry, Faculty of Chemistry and Chemical Technology, University of Ljubljana, 1000 Ljubljana, Slovenia.
authors.forename: San
authors.initials: S
authors.lastname: Hadži
authors.affiliation: Structural Biology Brussels, Department of Biotechnology, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium; Structural Biology Research Center, VIB, Pleinlaan 2, B-1050 Brussels, Belgium.
authors.forename: Sarah
authors.initials: S
authors.lastname: Haesaerts
authors.affiliation: Structural Biology Brussels, Department of Biotechnology, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium; Structural Biology Research Center, VIB, Pleinlaan 2, B-1050 Brussels, Belgium. Electronic address: remy.loris@vib-vub.be.
authors.forename: Remy
authors.initials: R
authors.lastname: Loris
authors.affiliation: Structural Biology Brussels, Department of Biotechnology, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium; Structural Biology Research Center, VIB, Pleinlaan 2, B-1050 Brussels, Belgium. Electronic address: agarciap@vub.ac.be.
authors.forename: Abel
authors.initials: A
authors.lastname: Garcia-Pino
mesh_terms: D019843:Archaeal Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D046912:Multiprotein Complexes / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D019843:Archaeal Proteins; D001426:Bacterial Proteins; D046912:Multiprotein Complexes
keywords: Persisters; Protein purification; Refolding protocol; Toxin–antitoxin