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Stepwise oxidations play key roles in the structural and functional regulations of DJ-1
- Title
- Stepwise oxidations play key roles in the structural and functional regulations of DJ-1
- Authors
- Song, In-Kang; Kim, Mi-Sun; Ferrell, James E., Jr.; Shin, Dong-Hae; Lee, Kong-Joo
- Ewha Authors
- 이공주; 신동해
- SCOPUS Author ID
- 이공주; 신동해
- Issue Date
- 2021
- Journal Title
- BIOCHEMICAL JOURNAL
- ISSN
- 0264-6021
1470-8728
- Citation
- BIOCHEMICAL JOURNAL vol. 478, no. 19, pp. 3505 - 3525
- Publisher
- PORTLAND PRESS LTD
- Indexed
- SCIE; SCOPUS
- Document Type
- Article
- Abstract
- DJ-1 is known to play neuroprotective roles by eliminating reactive oxygen species (ROS) as an antioxidant protein. However, the molecular mechanism of DJ-1 function has not been well elucidated. This study explored the structural and functional changes of DJ-1 in response to oxidative stress. Human DJ-1 has three cysteine residues (Cys46, Cys53 and Cys106). We found that, in addition to Cys106, Cys46 is the most reactive cysteine residue in DJ-1, which was identified employing an NPSB-B chemical probe (Ctag) that selectively reacts with redox-sensitive cysteine sulfhydryl. Peroxidatic Cys46 readily formed an intra-disulfide bond with adjacent resolving Cys53, which was identified with nanoUPLC-ESI-q-TOF tandem mass spectrometry (MS/MS) employing DBond algorithm under the non-reducing condition. Mutants (C46A and C53A), not forming Cys46-Cys53 disulfide cross-linking, increased oxidation of Cys106 to sulfinic and sulfonic acids. Furthermore, we found that DJ-1 C46A mutant has distorted unstable structure identified by biochemical assay and employing hydrogen/deuterium exchange-mass spectrometry (HDX-MS) analysis. All three Cys mutants lost antioxidant activities in SN4741 cell, a dopaminergic neuronal cell, unlike WT DJ-1. These findings suggest that all three Cys residues including Cys46-Cys53 disulfide cross-linking are required for maintaining the structural integrity, the regulation process and cellular function as an antioxidant protein. These studies broaden the understanding of regulatory mechanisms of DJ-1 that operate under oxidative conditions.
- DOI
- 10.1042/BCJ20210245
- Appears in Collections:
- 약학대학 > 약학과 > Journal papers
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