Site-specific and redox-controlled S-nitrosation of thioredoxin
Protein S-nitrosation on cysteine residues has emerged as an important posttranslational modification in mammalian cells. Previous studies have suggested a primary role for thioredoxin (Trx) in controlling protein S-nitrosation reactions. Human Trx contains five conserved Cys, including two redox-ac...
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| Format: | Article |
| Language: | en_US |
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National Academy of Sciences (U.S.)
2012
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| Online Access: | http://hdl.handle.net/1721.1/70140 |
| _version_ | 1826217415539687424 |
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| author | Barglow, Katherine T. Knutson, Charles G. F. Wishnok, John S. Tannenbaum, Steven Robert Marletta, Michael A. |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Barglow, Katherine T. Knutson, Charles G. F. Wishnok, John S. Tannenbaum, Steven Robert Marletta, Michael A. |
| author_sort | Barglow, Katherine T. |
| collection | MIT |
| description | Protein S-nitrosation on cysteine residues has emerged as an important posttranslational modification in mammalian cells. Previous studies have suggested a primary role for thioredoxin (Trx) in controlling protein S-nitrosation reactions. Human Trx contains five conserved Cys, including two redox-active catalytic Cys (Cys32 and Cys35) and three non-active-site Cys (Cys62, Cys69, and Cys73), all of which have been reported as targets of S-nitrosation. Prior reports have studied thermodynamic end points of nitrosation reactions; however, the kinetics of Trx nitrosation has not previously been investigated. Using the transnitrosation agent, S-nitrosoglutathione, a kinetic analysis of the selectivity and redox dependence of Trx nitrosation at physiologically relevant concentrations and times was performed, utilizing a mass spectrometry-based method for the direct analysis of the nitrosated Trx. Reduced Trx (rTrx) was nitrosated 2.7-times faster than oxidized Trx (oTrx), and rTrx was nitrosated selectively on Cys62, whereas oTrx was nitrosated only on Cys73. These sites of nitrosation were confirmed at the peptide level using a novel modification of the biotin-switch technique called the reductive switch. These results suggest separate signaling pathways for Trx-SNO under different cellular redox states. |
| first_indexed | 2024-09-23T17:03:17Z |
| format | Article |
| id | mit-1721.1/70140 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T17:03:17Z |
| publishDate | 2012 |
| publisher | National Academy of Sciences (U.S.) |
| record_format | dspace |
| spelling | mit-1721.1/701402022-10-03T10:04:30Z Site-specific and redox-controlled S-nitrosation of thioredoxin Barglow, Katherine T. Knutson, Charles G. F. Wishnok, John S. Tannenbaum, Steven Robert Marletta, Michael A. Massachusetts Institute of Technology. Department of Biological Engineering Tannenbaum, Steven Robert Tannenbaum, Steven Robert Knutson, Charles G. F. Protein S-nitrosation on cysteine residues has emerged as an important posttranslational modification in mammalian cells. Previous studies have suggested a primary role for thioredoxin (Trx) in controlling protein S-nitrosation reactions. Human Trx contains five conserved Cys, including two redox-active catalytic Cys (Cys32 and Cys35) and three non-active-site Cys (Cys62, Cys69, and Cys73), all of which have been reported as targets of S-nitrosation. Prior reports have studied thermodynamic end points of nitrosation reactions; however, the kinetics of Trx nitrosation has not previously been investigated. Using the transnitrosation agent, S-nitrosoglutathione, a kinetic analysis of the selectivity and redox dependence of Trx nitrosation at physiologically relevant concentrations and times was performed, utilizing a mass spectrometry-based method for the direct analysis of the nitrosated Trx. Reduced Trx (rTrx) was nitrosated 2.7-times faster than oxidized Trx (oTrx), and rTrx was nitrosated selectively on Cys62, whereas oTrx was nitrosated only on Cys73. These sites of nitrosation were confirmed at the peptide level using a novel modification of the biotin-switch technique called the reductive switch. These results suggest separate signaling pathways for Trx-SNO under different cellular redox states. American Cancer Society (Postdoctoral Fellowship) National Institutes of Health (U.S.) (NIH CA26731) Massachusetts Institute of Technology. Center for Environmental Health Sciences (ES002109) National Institutes of Health (U.S.) (NIH GM080272) 2012-04-25T20:59:25Z 2012-04-25T20:59:25Z 2011-08 2011-06 Article http://purl.org/eprint/type/JournalArticle 0027-8424 1091-6490 http://hdl.handle.net/1721.1/70140 Barglow, K. T. et al. “Site-specific and Redox-controlled S-nitrosation of Thioredoxin.” Proceedings of the National Academy of Sciences 108.35 (2011): E600–E606. Web. en_US http://dx.doi.org/10.1073/pnas.1110736108 Proceedings of the National Academy of Sciences of the United States of America Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf National Academy of Sciences (U.S.) PNAS |
| spellingShingle | Barglow, Katherine T. Knutson, Charles G. F. Wishnok, John S. Tannenbaum, Steven Robert Marletta, Michael A. Site-specific and redox-controlled S-nitrosation of thioredoxin |
| title | Site-specific and redox-controlled S-nitrosation of thioredoxin |
| title_full | Site-specific and redox-controlled S-nitrosation of thioredoxin |
| title_fullStr | Site-specific and redox-controlled S-nitrosation of thioredoxin |
| title_full_unstemmed | Site-specific and redox-controlled S-nitrosation of thioredoxin |
| title_short | Site-specific and redox-controlled S-nitrosation of thioredoxin |
| title_sort | site specific and redox controlled s nitrosation of thioredoxin |
| url | http://hdl.handle.net/1721.1/70140 |
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