Oxidative inactivation of thioredoxin as a cellular growth factor and protection by a Cys73 → Ser mutation

Gasdaska, John R.; Kirkpatrick, David; Montfort, W.R.; Kuperus, Miles; Hill, Simon; Berggren, Margareta; Powis, Garth

doi:10.1016/s0006-2952(96)00595-3

Cited by 65 publications

(54 citation statements)

References 31 publications

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“…Therefore, it is more likely that an oxidative modification of the protein occurs in the culture medium. This hypothesis is reinforced by the fact that the extracellular environment has been described to be predominantly oxidizing (61). The effect of rhTrx-1 in our study is probably mediated by a cell surface molecule and not following its infiltration to the cytoplasm (55), where it is less likely to exist under the oxidized form since the interior of the cell is highly reducing (61).…”

Section: Discussionsupporting

confidence: 49%

Extracellular Human Thioredoxin-1 Inhibits Lipopolysaccharide-induced Interleukin-1β Expression in Human Monocyte-derived Macrophages

Billiet

Furman

Larigauderie

et al. 2005

Journal of Biological Chemistry

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Oxidative stress plays an important role in atherosclerotic vascular disease, and several recent studies were focused on thioredoxin-1 (Trx-1) and its potential protective role against oxidative stress. Since human monocyte-derived macrophages (HMDM) are important cells in several inflammatory diseases including atherosclerosis, we conducted this study to evaluate the impact of extracellular recombinant human Trx-1 (rhTrx-1) on gene expression in lipopolysaccharide-activated HMDM. Our results showed that rhTrx-1 was capable of reducing interleukin (IL)-1␤ mRNA and protein synthesis in a dose-dependent manner. This effect was partly mediated through a reduction of NF-B activation as analyzed by transient transfection and gel shift assays. In addition, we showed that the attenuation of NF-B activity was the result of the reduction of both p50 and p65 subunit mRNA and protein synthesis on one hand and of the induction of I-B␣ mRNA and protein expression on the other hand. Moreover, inhibition of endogenous Trx-1 mRNA was also observed, suggesting a contribution to the diminution of NF-B activity since endogenous Trx-1, in contrast to the exogenous Trx-1, activates the NF-B system. Finally, H 2 O 2 -oxidized rhTrx-1 reduced IL-1␤ mRNA synthesis in lipopolysaccharide-activated HMDM. This result highly suggested that the rhTrx-1 used in this study could be oxidized in the culture medium and, in turn, reduced IL-1␤ mRNA and protein synthesis. Taken together, these data indicated a potential new mechanism through which extracellular rhTrx-1 exerts an anti-inflammatory function in HMDM.Increasing evidence indicates that oxidative stress plays an important role in atherosclerotic vascular disease (1). However, the protective mechanisms of antioxidants, including dietary antioxidants such as ascorbate (vitamin C) and ␣-tocopherol (vitamin E), have not been well defined (1). Nevertheless, recent attention was focused on thioredoxin-1 (Trx-1) 2 and its potential protective role against oxidative stress (see review (2)). Indeed, Trx-1, a 12-kDa highly conserved protein in almost all species, is the major carrier of redox potential in cells (3). Trx-1 functions by the reversible oxidization of two Trx-specific redoxactive cysteine residues (Cys-32 and Cys-35) to form a disulfide bond that, in turn, can be reduced by the action of Trx-1 reductase and NADPH.Intracellular Trx-1 exerts most of its antioxidant properties through scavenging of reactive oxygen species (4). In addition, it acts as a cofactor for several enzymes such as nucleoside diphosphate reductase (5), T7 DNA polymerase (6), and 3Јphosphoadenosylsulfate reductase (7). Moreover, mammalian Trx-1 plays an important role in the regulation of redox-sensitive transcription factors such as activator protein-1 (AP-1) and nuclear factor-B (NF-B) (8, 9) and in the regulation of glucocorticoid receptor-mediated signal transduction (9 -11).In addition to its intracellular role, intact Trx-1 and its truncated form (Trx-80) can be released by cells and exert both cytokine-like ...

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Section: Discussionsupporting

confidence: 49%

Extracellular Human Thioredoxin-1 Inhibits Lipopolysaccharide-induced Interleukin-1β Expression in Human Monocyte-derived Macrophages

Billiet

Furman

Larigauderie

et al. 2005

Journal of Biological Chemistry

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“…Glutathionylation has been implicated in buffering of oxidative stress, regulation of enzyme activity, and protection against potentially irreversible protein oxidation or polymerization (4,42,43). For example, H 2 O 2 -or diamide-treated thioredoxin is converted into an enzymatically inactive homodimer via a Cys 72 -Cys 72 intermolecular disulfide bond (44). Glutathionylation of the Cys 72 residue of thioredoxin prevents dimerization and is reversible (45).…”

Section: Discussionmentioning

confidence: 99%

Protein Disulfide Bond Formation in the Cytoplasm during Oxidative Stress

Cumming

Andon

Haynes

et al. 2004

Journal of Biological Chemistry

415

358

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The majority of disulfide-linked cytosolic proteins are thought to be enzymes that transiently form disulfide bonds while catalyzing oxidation-reduction (redox) processes. Recent evidence indicates that reactive oxygen species can act as signaling molecules by promoting the formation of disulfide bonds within or between select redox-sensitive proteins. However, few studies have attempted to examine global changes in disulfide bond formation following reactive oxygen species exposure. Here we isolate and identify disulfide-bonded proteins (DSBP) in a mammalian neuronal cell line (HT22) exposed to various oxidative insults by sequential nonreducing/reducing two-dimensional SDS-PAGE combined with mass spectrometry. By using this strategy, several known cytosolic DSBP, such as peroxiredoxins, thioredoxin reductase, nucleoside-diphosphate kinase, and ribonucleotide-diphosphate reductase, were identified. Unexpectedly, a large number of previously unknown DSBP were also found, including those involved in molecular chaperoning, translation, glycolysis, cytoskeletal structure, cell growth, and signal transduction. Treatment of cells with a wide range of hydrogen peroxide concentrations either promoted or inhibited disulfide bonding of select DSBP in a concentration-dependent manner. Decreasing the ratio of reduced to oxidized glutathione also promoted select disulfide bond formation within proteins from cytoplasmic extracts. In addition, an epitope-tagged version of the molecular chaperone HSP70 forms mixed disulfides with both ␤4-spectrin and adenomatous polyposis coli protein in the cytosol. Our findings indicate that disulfide bond formation within families of cytoplasmic proteins is dependent on the nature of the oxidative insult and may provide a common mechanism used to control multiple physiological processes.Oxidative stress occurs when the rate of reactive oxygen species (ROS) 1 generation exceeds the detoxification abilities of the cell, and it has been implicated in many degenerative diseases. It is frequently argued that ROS cause relatively nonspecific damage to vital cellular components such as lipids, DNA, and proteins. However, emerging evidence indicates that ROS can cause specific protein modifications that may lead to a change in the activity or function of the oxidized protein (1, 2). Several major forms of oxidative modifications can occur on amino acid residue side chains including carbonylation, nitrosylation, and oxidation of methionine to methionine sulfoxide (3). Protein sulfhydryls can be oxidized to protein disulfides and sulfenic acids as well as more highly oxidized states such as the sulfinic and sulfonic acid forms of protein cysteines (4). Under non-stressed conditions, disulfide bond formation occurs primarily in the oxidizing environment of the endoplasmic reticulum (ER) in eukaryotic cells (5). The sulfhydryl groups in the vast majority of protein cysteine residues (Cys-SH) have a pK a Ͼ8.0 and, in the reducing environment of the cytoplasm, remain protonated at physiological pH. Thus,...

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“…4) underlining the role of these residues for Trx binding and catalysis. In this work we used a C73S hTrx1 mutant in order to avoid formation of hTrx1 dimers involving a disulphide between Cys73 hTrx1 and C73′ hTrx1 of a second hTrx1 molecule 26,27 . Such Trx1-dimers are known not to be a substrate of TrxR1 (ref.…”

Section: Figure 6 | Close-up View Of the Intermolecular Disulphide Bondmentioning

confidence: 99%

“…Such Trx1-dimers are known not to be a substrate of TrxR1 (ref. 26). This finding can be explained by our structure, because in the presence of a disulphide between Cys73 hTrx1 and Cys73′ hTrx1 , the hTrx1 residues 73 hTrx1 to 75 hTrx1 , which are involved in binding of hTrx1 to hTrxR1 (see above), are not available for hTrxR1-hTrx1 complex formation (Fig.…”

Section: Figure 6 | Close-up View Of the Intermolecular Disulphide Bondmentioning

confidence: 99%

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Crystal structure of the human thioredoxin reductase–thioredoxin complex

et al. 2011

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Oxidative inactivation of thioredoxin as a cellular growth factor and protection by a Cys73 → Ser mutation

Cited by 65 publications

References 31 publications

Extracellular Human Thioredoxin-1 Inhibits Lipopolysaccharide-induced Interleukin-1β Expression in Human Monocyte-derived Macrophages

Extracellular Human Thioredoxin-1 Inhibits Lipopolysaccharide-induced Interleukin-1β Expression in Human Monocyte-derived Macrophages

Protein Disulfide Bond Formation in the Cytoplasm during Oxidative Stress

Crystal structure of the human thioredoxin reductase–thioredoxin complex

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