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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-KangKim, Mi-SunFerrell, James E., Jr.Shin, Dong-HaeLee, Kong-Joo
Ewha Authors
이공주신동해
SCOPUS Author ID
이공주scopusscopus; 신동해scopus
Issue Date
2021
Journal Title
BIOCHEMICAL JOURNAL
ISSN
0264-6021JCR Link

1470-8728JCR Link
Citation
BIOCHEMICAL JOURNAL vol. 478, no. 19, pp. 3505 - 3525
Publisher
PORTLAND PRESS LTD
Indexed
SCIE; SCOPUS WOS 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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