Stepwise oxidations play key roles in the structural and functional regulations of DJ-1
- 주제(기타) Biochemistry & Molecular Biology
- 설명문(일반) [Song, In-Kang; Kim, Mi-Sun; Shin, Dong-Hae; Lee, Kong-Joo] Ewha Womans Univ, Coll Pharm, Seoul 03760, South Korea; [Song, In-Kang; Kim, Mi-Sun; Shin, Dong-Hae; Lee, Kong-Joo] Ewha Womans Univ, Grad Sch Pharmaceut Sci, Seoul 03760, South Korea; [Ferrell, James E., Jr.] Stanford Univ, Dept Chem & Syst Biol, Sch Med, Stanford, CA 94305 USA
- 등재 SCIE, SCOPUS
- OA유형 hybrid, Green Accepted, Green Submitted
- 발행기관 PORTLAND PRESS LTD
- 발행년도 2021
- 총서유형 Journal
- URI http://www.dcollection.net/handler/ewha/000000183675
- 본문언어 영어
- Published As http://dx.doi.org/10.1042/BCJ20210245
- PubMed https://pubmed.ncbi.nlm.nih.gov/34515295
초록/요약
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.
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