Regulation of the Catalytic Activity and Structure of Human Thioredoxin 1 via Oxidation and S-Nitrosylation of Cysteine Residues
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The mammalian cytosolic/nuclear thioredoxin system, comprising thioredoxin (Trx), selenoenzyme thioredoxin reductase (TrxR), and NADPH, is the major protein-disulfide reductase of the cell and has numerous functions. The active site of reduced Trx comprises Cys(32)-Gly-Pro-Cys(35) thiols that catalyze target disulfide reduction, generating a disulfide. Human Trx1 has also three structural Cys residues in positions 62, 69, and 73 that upon diamide oxidation induce a second Cys(62)-Cys(69) disulfide as well as dimers and multimers. We have discovered that after incubation with H(2)O(2) only monomeric two-disulfide molecules are generated, and they are inactive but able to regain full activity in an autocatalytic process in the presence of NADPH and TrxR. There are conflicting results regarding the effects of S-nitrosylation on Trx antioxidant functions and which residues are involved. We found that S-nitrosoglutathione-mediated S-nitrosylation at physiological pH is critically dependent on the redox state of Trx. Starting from fully reduced human Trx, both Cys(69) and Cys(73) were nitrosylated, and the active site formed a disulfide; the nitrosylated Trx was not a substrate for TrxR but regained activity after a lag phase consistent with autoactivation. Treatment of a two-disulfide form of Trx1 with S-nitrosoglutathione resulted in nitrosylation of Cys(73), which can act as a trans-nitrosylating agent as observed by others to control caspase 3 activity (Mitchell, D. A., and Marletta, M. A. (2005) Nat. Chem. Biol. 1, 154-158). The reversible inhibition of human Trx1 activity by H(2)O(2) and NO donors is suggested to act in cell signaling via temporal control of reduction for the transmission of oxidative and/or nitrosative signals in thiol redox control.
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The Trx and Grx systems are required for the activity of ribonucleotide reductase (RNR), which provides deoxyribonucleotides for DNA replication and repair.
“These systems are present in all mammalian cells19,20, and are required for the activity of ribonucleotide reductase (RNR) which provides deoxyribonucleotides for DNA replication and repair.”
From Inhibition of the glutaredoxin and thioredoxin systems and ribonucleotide reductase by mutant p53-targeting compound APR-246 (2018) · cites this paper as [20]
The Trx and Grx systems are important antioxidants maintaining cellular redox balance and providing electrons for thiol-dependent reactions.
“The thioredoxin (Trx) and glutaredoxin (Grx) systems are important as antioxidants for maintaining cellular redox balance and providing electrons for thiol-dependent reactions like those catalyzed by ribonucleotide reductase and peroxiredoxins (Prxs).”
From Inhibition of the glutaredoxin and thioredoxin systems and ribonucleotide reductase by mutant p53-targeting compound APR-246 (2018) · cites this paper as [20]
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- Authors
- Hashemy, S. I., & Holmgren, A.
- Affiliations
- Karolinska Institutet (SE)
- Corresponding author
- Holmgren, Arne
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2008-08
- Volume
- 283
- Issue
- 32
- Pages
- 21890-21898
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W1993780555
- Cited by
- 192
- References
- 60
- Citations by year
- 2026: 3 · 2025: 9 · 2024: 6 · 2023: 6 · 2022: 11 · 2021: 14 · 2020: 11 · 2019: 6 · 2018: 13 · 2017: 14 · 2016: 10 · 2015: 9 · 2014: 15 · 2013: 9 · 2012: 15
- FWCI
- 5.60
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glutathione Transferases and Polymorphisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Sulfur Compounds in Biology
- Keywords
- thioredoxin-1, S-nitrosylation, S-nitrosoglutathione, cysteine residues, disulfide bond formation, thioredoxin reductase, protein disulfide isomerase, self-activation, hydrogen peroxide, nitric oxide donors, protein dimerization, caspase-3, antioxidant activity
- MeSH terms
- Catalysis, Cysteine, Diamide, Humans, Hydrogen Peroxide, Models, Molecular, Oxidation-Reduction, Proline, Sulfhydryl Reagents, Thioredoxins, Time Factors, Oxidants, Protein Structure, Tertiary, Nitric Oxide Donors, S-Nitrosoglutathione
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