Purification of Recombinant Human PARG and Activity Assays
| Autor: | Jean-Christophe, Amé, Éléa, Héberlé, Barbara, Camuzeaux, Françoise, Dantzer, Valérie, Schreiber |
|---|---|
| Přispěvatelé: | Biotechnologie et signalisation cellulaire (BSC), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut de recherche de l'Ecole de biotechnologie de Strasbourg (IREBS) |
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
Poly Adenosine Diphosphate Ribose
PARG purification MESH: Humans Glycoside Hydrolases MESH: Escherichia coli enzymology Sciences du Vivant [q-bio]/Biotechnologies MESH: Biological Assay Recombinant Proteins MESH: Recombinant Proteins isolation & purification metabolism MESH: Poly Adenosine Diphosphate Ribose MESH: Protein Processing Post-Translational Escherichia coli MESH: Glycoside Hydrolases Animals Humans Biological Assay MESH: Animals [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Poly(ADP-ribose) glycohydrolase Protein Processing Post-Translational Poly(ADP-ribose) metabolism |
| Zdroj: | Methods Mol Biol Methods Mol Biol, 2017, 1608, pp.395-413. ⟨10.1007/978-1-4939-6993-7_25⟩ |
| DOI: | 10.1007/978-1-4939-6993-7_25⟩ |
| Popis: | The purification of Poly(ADP-ribose) glycohydrolase (PARG) from overexpressing bacteria Escherichia coli is described here to a fast and reproducible one chromatographic step protocol. After cell lysis, GST-PARG-fusion proteins from the crude extract are affinity purified by a Glutathione 4B Sepharose chromatographic step. The PARG proteins are then freed from their GST-fusion by overnight enzymatic cleavage using the preScission protease. As described in the protocol, more than 500 μg of highly active human PARG can be obtained from 1.5 L of E. coli culture. journal article 2017 imported |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|