Purification of Recombinant Human PARG and Activity Assays.
Fiche publication
Date publication
janvier 2023
Journal
Methods in molecular biology (Clifton, N.J.)
Auteurs
Membres identifiés du Cancéropôle Est :
Dr DANTZER Françoise
Tous les auteurs :
Amé JC, Dantzer F
Lien Pubmed
Résumé
The purification of poly(ADP-ribose) glycohydrolase (PARG) from overexpressing bacteria Escherichia coli is described here as 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.
Mots clés
PARG purification, Poly(ADP-ribose) glycohydrolase, Poly(ADP-ribose) metabolism
Référence
Methods Mol Biol. 2023 ;2609:399-418