Abstract
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.
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Amé, J. C., Héberlé, É., Camuzeaux, B., Dantzer, F., & Schreiber, V. (2017). Purification of recombinant human PARG and activity assays. In Methods in Molecular Biology (Vol. 1608, pp. 395–413). Humana Press Inc. https://doi.org/10.1007/978-1-4939-6993-7_25
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