| Autor: |
Yann Sterckx, STEVEN DE GIETER, Valentina Zorzini, San Hadzi, Sarah Haesaerts, Remy Loris, Abel Garcia Pino |
| Přispěvatelé: |
Structural Biology Brussels, Department of Bio-engineering Sciences |
| Předmět: |
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| Zdroj: |
Vrije Universiteit Brussel |
| 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: |
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