University of Oulu

Jiro Ogura, Lloyd W. Ruddock, Nariyasu Mano, Cysteine 343 in the substrate binding domain is the primary S-Nitrosylated site in protein disulfide isomerase, Free Radical Biology and Medicine, Volume 160, 2020, Pages 103-110, ISSN 0891-5849, https://doi.org/10.1016/j.freeradbiomed.2020.07.029

Cysteine 343 in the substrate binding domain is the primary S-Nitrosylated site in protein disulfide isomerase

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Author: Ogura, Jiro1,2; Ruddock, Lloyd W.2; Mano, Nariyasu1
Organizations: 1Department of Pharmaceutical Sciences, Tohoku University Hospital, Sendai, Japan
2Faculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu, Finland
Format: article
Version: accepted version
Access: embargoed
Persistent link: http://urn.fi/urn:nbn:fi-fe202103056643
Language: English
Published: Elsevier, 2020
Publish Date: 2021-08-05
Description:

Abstract

Abnormal protein accumulations are typical pathological features for neurodegenerative diseases. Protein disulfide isomerase (PDI) is a critical enzyme in oxidative protein folding. S-nitrosylated PDI has been detected in the postmortem brain in neurodegenerative disease patients, but the effect of S-nitrosylation on PDI function and developing neurodegeneration was not clarified in detail. In this study, we identified that in vitro and in vivo S-nitrosylation of C343 in the b’ domain of PDI occurs. Reduced recombinant human PDI (hPDI) reacted quickly with S-nitrosocompounds, with an observed increase in the expected S-nitrosylated species and the appearance of the disulfide state of the active sites. Both Mononitrosylated and dinitrosylated were observed from the S-nitrosylation of hPDI. Dinitrosylated species were S-nitrosylated both cysteines at active site. But, at least in part, mononitrosylated species were S-nitrosylated on cysteine 343 in the substrate binding b’ domain. Although active site S-nitrosylation is reversible by reduced glutathione, S-nitrosylation of C343 is comparative stable. S-nitrosylation of PDI in SH-SY5Y cells was observed after the S-nitrosocysteine (SNOC) treatment and S-nitrosylated PDI was still detected 24 h after removing SNOC. While wild-type PDI was S-nitrosylated, the level of S-nitrosylation of the C343S mutant in over-expressed cells was substantially lower and only wild-type PDI of S-nitrosylation remained 24 h after removing SNOC in over-expressed cells. Both of in vitro and in vivo results suggested that S-nitrosylation of C343 in PDI may be the causative effect on physiological changes in neurodegerenative disease patients, and may be useful for the drug development for neurodegenerative diseases.

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Series: Free radical biology & medicine
ISSN: 0891-5849
ISSN-E: 1873-4596
ISSN-L: 0891-5849
Volume: 160
Pages: 103 - 110
DOI: 10.1016/j.freeradbiomed.2020.07.029
OADOI: https://oadoi.org/10.1016/j.freeradbiomed.2020.07.029
Type of Publication: A1 Journal article – refereed
Field of Science: 1182 Biochemistry, cell and molecular biology
Subjects:
Funding: This study was in part supported by the Uehara Memorial Foundation fellowships to J.O. and the Japan Society for the Promotion of Science (JSPS) KAKENHI grant number 18H06099 and 19K21219 to J.O.
Copyright information: © 2020 Elsevier B.V. This manuscript version is made available under the CC-BY-NC-ND 4.0 license by http://creativecommons.org/licenses/by-nc-nd/4.0/.
  https://creativecommons.org/licenses/by-nc-nd/4.0/