Separation of native and C106-oxidized DJ-1 proteins by using column chromatography

• Published in: Protein expression and purification Vol. 195-196; p. 106092
• Main Authors: Choi, Joonhyeok, Yoo, Hee-Jin, Cho, Kun, Kim, Hak Nam, Lee, Joon-Hwa, Ryu, Kyoung-Seok
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.08.2022
• Subjects: Chromatography; DJ-1; DJ-1 C106 sulfinic acid; DJ-1 oxidation; DJ-1 purification; Humans; Oncogene Proteins - chemistry; Oncogene Proteins - genetics; Oncogene Proteins - metabolism; Oxidation-Reduction; Oxidative Stress; Parkinson Disease - genetics; Parkinson Disease - metabolism; Parkinson's disease; Protein Deglycase DJ-1 - genetics; Protein Deglycase DJ-1 - metabolism; DJ-1 purification; Parkinson's disease; matrix-assisted laser desorption/ionization time of flight mass; size-exclusion column chromatography; transverse relaxation-optimized spectroscopy-heteronuclear single quantum coherence; DJ-1; DJ-1 oxidation; DJ-1 C106 sulfinic acid; Dithiothreitol; nuclear magnetic resonance; isoelectric focusing gel electrophoresis
• ISSN: 1046-5928; 1096-0279
• DOI: 10.1016/j.pep.2022.106092

Mutations in PARK7, the gene encoding the DJ-1 protein, are associated with early onset of Parkinson's disease. The C106 residue of DJ-1 is highly susceptible to oxidation, and its oxidation status is essential for various in vivo neuroprotective roles. Since C106 is readily oxidized to sulfinic acid that is not reduced by dithiothreitol, no method to separate native DJ-1 protein from the oxidized one creates challenges in the in vitro study of the biological relevance of C106-oxidation state. Here, we report an efficient column chromatography method to purify native, C106-sulfinic, and mixed (combination of the priors) forms of DJ-1. This method will be useful for systematic in vitro studies of DJ-1 functions by providing specific native and C106-sulfinic DJ-1 proteins. •DJ-1 is one of the key proteins involved in the early onset of Parkinson's disease.•C106 oxidation (e.g. sulfinic) is crucial for the neuroprotective functions of DJ-1.•Difficulty in the specific management of native and C106-oxidized DJ-1 proteins.•C106 of DJ-1 is highly susceptible to oxidation due to its low pKa value.•Cation-exchange column chromatography separates native and C106-sulfinic forms.