Published online before print December 13, 2007, doi: 10.1161/CIRCRESAHA.107.159814
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© 2008 American Heart Association, Inc.
Molecular Medicine |
Protein Carbonylation as a Novel Mechanism in Redox Signaling
From the Department of Pharmacology (C.M.W., Y.J.S.) and Lombardi Comprehensive Cancer Center (A.K.C., L.Z.), Georgetown University Medical Center, Washington, DC.
Correspondence to Dr Yuichiro J. Suzuki, Department of Pharmacology, Georgetown University Medical Center, NW403 Medical–Dental Building, 3900 Reservoir Rd NW, Washington, DC 20057. E-mail ys82{at}georgetown.edu
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Abstract |
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Reactive oxygen species serve as second messengers for signal transduction; however, molecular targets of oxidant signaling have not been defined. Here, we show that ligand–receptor–mediated signaling promotes reactive oxygen species–dependent protein carbonylation. Treatment of pulmonary artery smooth muscle cells with endothelin-1 increased protein carbonyls. Carbonylation of the majority of proteins occurred transiently, suggesting that there is also a mechanism for decarbonylation induced by endothelin-1. Decarbonylation was suppressed by inhibition of thioredoxin reductase, and cellular thioredoxin was upregulated during the decarbonylation phase. These results indicate that endothelin-1 promotes oxidant signaling as well as thioredoxin-mediated reductive signaling to regulate carbonylation and decarbonylation mechanisms. In cells treated with endothelin receptor antagonists, hydrogen peroxide scavengers, or an iron chelator, we identified, via mass spectrometry, proteins that are carbonylated in a receptor- and Fenton reaction–dependent manner, including annexin A1, which promotes apoptosis and suppresses cell growth. Carbonylation of annexin A1 by endothelin-1 was followed by proteasome-dependent degradation of this protein. We propose that carbonylation and subsequent degradation of annexin A1 may play a role in endothelin-mediated cell growth and survival, important events in pulmonary vascular remodeling. Protein carbonylation in response to ligand–receptor interactions represents a novel mechanism in redox signaling.
Key Words: endothelin-1 • protein carbonylation • oxidant signaling • pulmonary hypertension • smooth muscle
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Introduction |
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Reactive oxygen species (ROS) have been proposed to serve as second messengers for signal transduction processes.1–5 Numerous studies demonstrated that (1) ligand–receptor interactions produce ROS; (2) antioxidants block signal transduction; and (3) ROS can stimulate signaling events.6–11 ROS signaling is thought to play important roles in various diseases, including cancer, neurological disorders, immune diseases, and cardiovascular diseases. Although mechanisms of ROS actions during oxidant signaling have not been defined, protein thiols being the oxidation targets have been a popular concept.12–14 Recently, Lee and Helmann15 described a regulatory mechanism for the Bacillus subtilis PerR transcription factor by metal-catalyzed oxidation. Thus, other types of protein oxidation such as metal-catalyzed protein carbonylation may also be important for cell signaling.
Endothelin (ET)-1 is produced by vascular endothelial cells and exerts potent vasoconstrictive16 and mitogenic17 actions on vascular smooth muscle cells (SMCs). In pulmonary circulation, ET-1 contributes to vasoconstriction and vascular remodeling, which occur in pulmonary hypertension. The ET-1 expression is increased in the lungs of patients with pulmonary hypertension,18 and ET receptor antagonists have been used to treat human pulmonary hypertension,19,20 indicating the clinical importance of ET-1 signal transduction.
ET-1 is a mitogen of pulmonary artery SMCs21–23 via the activation of either ETA or ETB receptor.24 Signal transduction pathways induced by ET-1 in pulmonary artery SMCs, however, are not well understood. ET-1 can activate extracellular signal-regulated mitogen-activated protein kinase25 as well as GATA-4 transcription factor.26 ET-1 has also been shown to generate ROS, which promote pulmonary vascular SMC proliferation.27 Thus, ROS may play important roles in ET-1–mediated pulmonary artery SMC growth during the development of pulmonary hypertension.
The present study demonstrates that ET-1 promotes protein carbonylation in pulmonary artery SMCs in an ET receptor– and Fenton reaction–dependent fashion. We also identified the "decarbonylation" mechanism that is activated by ET-1 via thioredoxin. Proteins that are carbonylated in response to ET-1 were identified using 2D gel electrophoresis and mass spectrometry. Annexin A1, which inhibits cell growth, was found to be among the proteins that were carbonylated and subsequently degraded in response to ET-1 in a receptor and metal-catalyzed oxidation–dependent manner.
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Materials and Methods |
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Cell Culture
Large to midsized bovine pulmonary artery and thoracic aorta were obtained from a local abattoir. Bovine pulmonary artery SMCs (BPASMCs) and aortic SMCs were isolated as described previously28 and cultured in RPMI medium 1640 supplemented with 10% FBS, 1% penicillin/streptomycin, and 0.5% fungizone (Invitrogen, Carlsbad, Calif). Two to 8 passages were used for experiments. Cells were growth-arrested for 24 hours in medium supplemented with 0.01% FBS and then treated with ET-1 (Sigma-Aldrich, St Louis, Mo), H2O2 (Fisher Scientific, Hampton, NH), or 1-chloro-2,4-dinitrobenzene (DNCB) (Sigma-Aldrich) for 0, 5, 10, 15, 20, and 30 minutes. In some experiments, cells were pretreated for 30 minutes with BQ123, BQ788, ebselen, deferoxamine, MG132, or DNCB (Sigma-Aldrich).
Organ Culture
For organ culture of rat pulmonary artery, lungs, and hearts were obtained from male Sprague–Dawley rats (250 to 300 g) and placed in ice-cold PBS. Mid-to-large pulmonary arteries were dissected, and surrounding connective tissues were removed under a dissecting microscope. Blood vessels were cut into small pieces of ring segments and incubated in RPMI medium 1640 containing 0.01% FBS for 30 minutes at 37°C before treating with ET-1.
Statistical Analysis
Comparisons between 2 groups were analyzed by a 2-tailed Student’s t test, and comparisons among 3 or more groups were analyzed by ANOVA with a Student–Newman–Keuls post hoc test. P<0.05 was considered to be significant. Data are presented as means±SEM.
An expanded materials and methods section, as well as supplemental figures, can be found in the online data supplement at http://circres.ahajournals.org.
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Results |
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ET-1 Promotes Protein Carbonylation
Treatment of BPASMCs with ET-1 (30 nmol/L) resulted in the production of ROS, as monitored by detecting green dichlorofluorescein fluorescence (supplemental Figure I). The production of ROS was apparent after 5 minutes of ET-1 treatment, and the level of ET-1 that produced dichlorofluorescein fluorescence was comparable to the level produced by adding 50 µmol/L hydrogen peroxide (H2O2) extracellularly (supplemental Figure I). Thus, ET-1 produces ROS in BPASMCs.
Immunoblotting of 2,4-dinitrophenylhydrazine (DNPH)-derivatized proteins revealed that various proteins were carbonylated in BPASMCs (Figure 1A). Without DNPH derivatization, no bands were detected (data not shown). Treatment of cells with ET-1 for 10 minutes increased protein carbonylation of specific proteins, as determined by 1D (Figure 1A) and 2D gel electrophoresis (supplemental Figure IIA). Coomassie blue staining of the membranes showed no changes in total protein content (Figure 1A, bottom). We assigned numbers for each of carbonylated protein (CP) bands from 1D gel electrophoresis experiments to be CP1 to CP17 (Figure 1B). The ET-1–mediated promotion of carbonylation of these proteins was also found in the tissue culture of bovine pulmonary artery smooth muscle (supplemental Figure IIB) and in the organ culture of rat pulmonary artery (supplemental Figure IIC).
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Roles of ET Receptors
To determine whether ET-1–mediated protein carbonylation is dependent on ET receptors, BPASMCs were treated with BQ123 (ETA receptor antagonist)29 or BQ788 (ETB receptor antagonist)30 before treatment with ET-1. Results show that BQ123 inhibited ET-1–mediated carbonylation of CP8, -10, -11, -12, and -17 (Figure 2A and supplemental Figure IIIA); and BQ788 inhibited that of CP3, -5, -7, -10, -11, -12, -13, -15, and -17 (Figure 2B and supplemental Figure IIIB). Neither of the ET receptor antagonists influenced basal levels of protein carbonylation. This is the first demonstration of receptor-mediated signaling promoting protein carbonylation.
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Roles of H2O2 and Metal-Catalyzed Oxidation
CPs are often formed via peroxide-dependent Fenton reaction,31 and H2O2 has been shown to play integral roles in oxidant-mediated signal transduction.9,32 Consistently, pretreatment of BPASMCs with ebselen,33 a mimetic of glutathione peroxidase that decomposes H2O2, effectively inhibited all of the ET receptor–dependent protein carbonylation (Figure 2C and supplemental Figure IIIC). Similar results were obtained by catalase overexpression via adenovirus-mediated gene transfer (data not shown). Furthermore, H2O2 at the concentration as low as 0.5 µmol/L mimicked ET-1–mediated protein carbonylation (Figure 2D and supplemental Figure IIID). These results suggest that ET-1 promotes protein carbonylation via H2O2.
Recently, a transcription factor PerR was found to be regulated by metal-catalyzed oxidation.15 Thus, we tested the effects of deferoxamine, and we found that this iron chelator inhibited ET-1–mediated carbonylation of CP3, -7, -13, -15, and -17 (Figure 2E and supplemental Figure IIIE), suggesting the role of metal-catalyzed oxidation.
Kinetics of ET-1–Mediated Protein Carbonylation
As shown in Figure 1, the increases in protein carbonyls by ET-1 appear transient in cell culture of BPASMCs (Figure 1A) and in tissue culture of bovine pulmonary artery smooth muscle (supplemental Figure IIB). Analyses of individual bands further revealed that the levels of protein carbonyls were transiently increased. In BPASMCs, carbonylation of CP10, -11, -12, -13, and -15 increased by 5 minutes, peaked at 10 minutes, and decreased to the basal level by 30 minutes of ET-1 treatment (Figure 3A). In contrast, carbonylation of these CPs in bovine aortic SMCs increased rapidly by 5 minutes and was sustained for 20 to 30 minutes (Figure 3B), differing from transient kinetics as seen in SMCs of pulmonary circulation. These experiments identified that the ET-1–mediated stimulation of protein carbonylation is followed by normalization of CPs (we termed "decarbonylation") in pulmonary artery SMCs, whereas in aortic SMCs, such decarbonylation mechanism is slower.
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Mechanism of Decarbonylation
A decrease in CPs may be attributable to the proteasomal degradation of oxidatively modified proteins.34,35 To test this, BPASMCs were pretreated with MG132 (proteasome inhibitor) for 30 minutes before the ET-1 treatment. Figure 4 shows that ET-1–mediated increases in the majority of CPs occurred transiently even after the treatment with MG132 (30 µmol/L) for 30 minutes. MG132 can effectively inhibit proteolytic degradation in BPASMCs, because it inhibited the degradation of I
B-
induced by tumor necrosis factor- (supplemental Figure IV). Thus, the decarbonylation mechanism appears to be independent of the proteasomal degradation of oxidatively modified proteins.
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We hypothesized that reductive signaling may be promoted by ET-1, subsequently to oxidant signaling, perhaps to serve as a negative-feedback mechanism. Consistently, CPs in BPASMCs are reduced by β-mercaptoethanol (Figure 5). In these experiments, β-mercaptoethanol was added to BPASMC lysates before derivatization with DNPH. Also, an inhibitor of thioredoxin reductase, DNCB36 effectively promoted protein carbonylation (Figure 6A and supplemental Figure VA), suggesting the possible role of thioredoxin in reductive signaling. Unlike transient carbonylation induced by ET-1 in BPASMCs, DNCB-mediated carbonylation was sustained for at least 30 minutes without exhibiting decarbonylation. Treatment of BPASMCs with ET-1 after DNCB pretreatment further promoted some degree of protein carbonylation; however, ET-1 signaling did not elicit decarbonylation mechanism under these conditions (Figure 6B and supplemental Figure VB), demonstrating that the thioredoxin system is required for ET-1–mediated decarbonylation.
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Western blotting revealed that ET-1 time-dependently increased the levels of thioredoxin protein in BPASMCs (Figure 6C). Interestingly, not only did ET-1 fail to increase thioredoxin in aortic SMCs, but the basal level of aortic SMC thioredoxin was found to be substantially lower compared with that in pulmonary artery SMCs (Figure 6C), providing a possible mechanism for sustained kinetics of ET-1–mediated protein carbonylation and the lack of the decarbonylation mechanism in aortic SMCs. Actinomycin D, a general inhibitor of gene transcription, inhibited ET-1–mediated increase in thioredoxin expression (supplemental Figure VC), suggesting that ET-1 activates thioredoxin gene transcription. These results suggest that, in pulmonary artery SMCs, thioredoxin may play a role in ET-1 signal transduction and the regulation of decarbonylation.
To provide direct evidence that thioredoxin regulates protein carbonylation in pulmonary artery SMCs, the hypothesis that ET-1 promotes protein–protein interactions between thioredoxin and CPs was tested. Lysate samples of BPASMCs with or without ET-1 treatment were derivatized with DNPH and immunoprecipitated with the 2,4-dinitrophenylhydrazone (DNP) antibody, electrophoresed, and blotted with the thioredoxin antibody. Thioredoxin–CP interactions appear intact even in the presence of DNPH derivatization. Thioredoxin interactions with CP were found to be increased during the decarbonylation phase (ie, 20 to 30 minutes after ET-1 treatment) (Figure 6D). Thioredoxin itself is not carbonylated, because the thioredoxin band resides between CP16 and CP17 (data not shown). These results suggest that thioredoxin–carbonyl group interactions may regulate decarbonylation. Furthermore, surprisingly, we found that ET-1 treatment initially decreased thioredoxin–CP interactions during the period when CPs are increased (10 to 15 minutes), revealing the possibility that the mechanism of the promotion of carbonylation may involve the dissociation of thioredoxin from the target proteins. These results support experiments using DNCB, which provided evidence for the role of thioredoxin system in the regulation of decarbonylation.
Identification of Carbonylated Proteins
To determine the identities of proteins that are carbonylated in response to ET-1, we further analyzed 2D gels. Using information described in supplemental Figure IIA, in which the first dimension was separated with a pH range of 3 to 10 and 4% to 20% gradient gels were used for the second dimension, pH 6 to 8 isoelectric focusing with 10.5% to 14% gels were chosen for the amplification of the region with abundant ET-1–responsive CPs. We first identified 26 spots with intensities that were increased by ET-1 treatment for 10 minutes (supplemental Figure VIA). We then performed a series of experiments to refine the spots to ET-1–inducible CPs that were inhibitable by ET receptor antagonists ebselen and deferoxamine to define ET receptor– and Fenton reaction–dependent protein carbonylation. Supplemental Figure VIB shows that spot 10 is upregulated by ET-1, and this upregulation was attenuated by BQ123, BQ788, ebselen, and deferoxamine. All of the 26 spots were analyzed, and the results show that spots 8, 10, 11, 12, 13, 14, 16, 17, 20, 21, and 26 satisfied these criteria. The results of these experiments are shown in supplemental Figure VIB and VIC, and the locations of these 11 spots are summarized in supplemental Figure VID.
All of these 11 spots were excised from the gels and underwent mass spectrometric analyses. Mass spectrometric results are shown in supplemental Figure VII. Peptide mass fingerprinting and sequence analyses identified these proteins as heat shock protein β-1 (spot 8 and 16), peroxiredoxin 6 (spot 10 and 11), annexin A2 (spot 12 and 13), phosphoglycerate mutase 1 (spot 14), phosphoglycerate dehydrogenase (spot 17), cofilin-1 (spot 20), annexin A1 (spot 21), and DJ-1 protein (spot 26) with high confidence (>95%) and an average sequence coverage of 40%. We thus identified 8 proteins that are carbonylated in response to ET-1 in a receptor- and Fenton reaction–dependent manner, as summarized in Figure 7A.
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To confirm these mass spectrometric results, immunoprecipitation/immunoblot experiments were performed for peroxiredoxin 6 and annexin A1. Cell lysates from untreated and ET-1–treated cells were derivatized with DNPH, immunoprecipitated with DNP antibody, and blotted with either peroxiredoxin 6 (Figure 7B) or annexin A1 (Figure 7C). Similarly, carbonylation of annexin A2 and cofilin-1 were also confirmed (supplemental Figure VIII). The results demonstrate that carbonylation of these proteins are promoted by ET-1.
Consequences of Annexin A1 Carbonylation
Annexin A1 has been shown to inhibit proliferation and to promote apoptosis in various cell types.37–41 Similarly, in pulmonary artery SMCs, overexpression of annexin A1 reduced cell number and inhibited cell proliferation induced by platelet-derived growth factor and FBS (Figure 8A). We found that carbonylation of annexin A1 induced by ET-1 was followed by decreased annexin A1 protein expression (Figure 8B), whereas the protein expression of peroxiredoxin 6 was not altered (data not shown). The ET-1–mediated reduction of annexin A1 protein was inhibited by a proteasome inhibitor, MG132 (Figure 8C). These results suggest that, whereas many of proteins carbonylated in response to ET-1 signaling undergo decarbonylation, carbonylation of annexin A1 is followed by proteasome-dependent degradation. The degradation of annexin A1 should alleviate the actions of this protein to inhibit cell proliferation and promote apoptosis, and these events may be involved in the mechanism of ET-1–mediated pulmonary artery SMC growth.
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Discussion |
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In summary, here, we describe novel redox signaling events that occur in response to ligand–receptor interactions. In pulmonary artery SMCs, ET-1 promotes protein carbonylation via an ET receptor–dependent mechanism, followed by the reduction of CPs. We demonstrate that receptor-dependent ET-1 signaling for protein carbonylation is mediated by H2O2- and iron-dependent Fenton reaction. We also show that the subsequent reductive event termed "decarbonylation" appears to involve thioredoxin and thioredoxin reductase. Although the majority of proteins that are carbonylated in response to ET-1 undergo reduction-dependent decarbonylation, some CPs are degraded by proteasomes. Annexin A1 was found to be carbonylated by ET-1 in a receptor- and Fenton reaction–dependent manner and becomes degraded by proteasomes. Because annexin A1 functions to promote apoptosis and inhibit cell growth, the carbonylation-driven degradation of this protein may serve as mechanisms for cell growth and survival signaling by ET-1. These results show that ligand–receptor interactions can promote protein carbonylation events, which may play important functional roles.
Protein Carbonylation As a Novel Mechanism in Redox Signaling
The field of ROS had undergone a paradigm shift when the idea had emerged that these species are not merely damage-causing but can also serve as second messengers for signal transduction.1,2,4 Subsequent to the reports by Herzenberg and colleagues showing that antioxidants such as N-acetylcysteine can inhibit nuclear factor B and HIV activation,42,43 Baeuerle and colleagues proposed that H2O2 is a widely used second messenger for nuclear factor B activation in T cells.8 In vascular smooth muscle, early work demonstrated that ROS promoted cell growth, protooncogene expression,44 and Ca2+ signaling.10 Subsequently, ROS were reported to mediate signal transduction induced by angiotensin II6 and platelet-derived growth factor9 in aortic SMCs. In pulmonary artery SMCs, Fanburg and colleagues found that serotonin activates the production of superoxide45,46 and H2O2,47 presumably via NAD(P)H oxidase. We have shown that GATA-4 transcription factor plays an important role in pulmonary artery SMC growth and that antioxidants can inhibit serotonin-induced GATA-4 activation.26 Lawrie et al48 reported that, in human pulmonary artery SMCs, serotonin activates GATA-4 via ROS produced by monoamine oxidase. ET-1 has also been shown to produce ROS in fetal sheep pulmonary artery SMCs via NAD(P)H oxidase, and antioxidants block ET-1–induced cell proliferation,27 suggesting that ROS may play a role in ET-1–mediated pulmonary vascular thickening. The targets of ROS produced by ET-1 as well as other mediators of pulmonary hypertension, however, have not been defined. For the mechanisms of ROS signaling, ligand–receptor interactions producing ROS via NAD(P)H oxidase6,49 and ROS targeting protein thiols12,14 are currently popular proposed mechanisms.
During various oxidative stress conditions, protein oxidation results in the inactivation of protein functions. Carbonylation is one of oxidation processes, which can occur on protein molecules. Protein carbonyls are quite stable products formed on proline, arginine, lysine or threonine residues, often in response to metal-catalyzed Fenton reaction.31 Protein carbonyl groups react with DNPH, and various techniques have been developed to detect their interactions; these groups also have been used effectively as markers of oxidative stress. To our knowledge, the present study is the first demonstration of the protein carbonyl formation in response to ligand–receptor interactions and, therefore, the first demonstration of such formation in signal transduction processes. Protein carbonylation could offer a specific targeting mechanism for oxidant-mediated signal transduction, because site-directed Fenton reaction may occur at certain metal-binding sites within protein molecules.31 Further understanding of the roles of protein carbonylation in signal transduction pathways may yield invaluable information for the identification of targeting mechanisms for ROS signaling.
Discovery of the Decarbonylation Mechanism
It was interesting to note that carbonylation of the majority of the proteins were formed transiently in response to ET-1 in pulmonary artery SMCs. This led us to discover a process in which carbonylation can be eliminated in the cell. We termed this process "decarbonylation." Apparent decarbonylation could be observed if CPs were degraded, because oxidized proteins have been shown to be susceptible to proteolytic degradation.35 However, our experiments show that decarbonylation occurs even when proteasomes are inhibited, suggesting that the degradation mechanism may not explain our observations. It is plausible that CPs may be aggregated; however, because our cell lysis solution contains detergent, it is expected that these proteins remain in the supernatant after centrifugation. We propose that decarbonylation is dependent on reduction reactions through several lines of evidence. First, CPs are sensitive to and can be eliminated by reactions with reductants such as β-mercaptoethanol. Second, a thioredoxin reductase inhibitor can promote protein carbonylation without the occurrence of decarbonylation. Thirdly, when thioredoxin reductase is inhibited, ET-1 promotes protein carbonylation in a sustained fashion without the occurrence of decarbonylation. Also, it is interesting to note that thioredoxin is upregulated in pulmonary artery SMCs in response to ET-1, and its expression is substantially lower in aortic SMCs, where the decarbonylation event is not apparent. These results indicate that thioredoxin, which has been shown to play integral roles in redox signaling,50 may also regulate signal transduction mechanism involving protein carbonylation and decarbonylation. Because protein carbonylation is not chemically reversible,51 the nature of decarbonylation is not yet known.
Clinical Implications
Because ET receptor antagonists are used for the treatment of human pulmonary hypertension, these redox regulatory mechanisms promoted by ET-1 may offer important insights to therapeutic strategies. Our laboratory has previously demonstrated that ET-1 can promote anti-apoptotic signaling in pulmonary vascular SMCs,52 suggesting that ET-1 may exert multiple actions to contribute to the development of pulmonary hypertension. Because proteins that were found to be carbonylated in response to ET-1 signaling include annexin A1, which has been shown to promote apoptosis and inhibit cell proliferation,37–41 protein carbonylation of this protein by ET-1 may increase cell growth. In fact, we found that ET-1–mediated carbonylation of annexin A1 was followed by proteasome-dependent degradation, consistent with the idea that ET-1 alleviates apoptotic and antiproliferative actions of annexin A1 for promoting survival signaling and increasing SMC number. Further studies of carbonylated annexin A1, as well as other identified proteins, should provide important information for determining the mechanism of redox regulation of signal transduction, as well as for identifying effective therapeutic interventions against pulmonary hypertension.
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Acknowledgments |
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Sources of Funding
This work was supported in part by NIH grants HL067340 and HL072844 (to Y.J.S.).
Disclosures
None.
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Footnotes |
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Original received July 12, 2007; revision received November 1, 2007; accepted November 29, 2007.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
1. Archer SL, Huang J, Henry T, Peterson D, Weir EK. A redox-based O2 sensor in rat pulmonary vasculature. Circ Res. 1993; 73: 1100–1112.
2. Weir EK, Archer SL. The mechanism of acute hypoxic pulmonary vasoconstriction: the tale of two channels. FASEB J. 1995; 9: 183–189.[Abstract]
3. Sen CK, Packer L. Antioxidant and redox regulation of gene transcription. FASEB J. 1996; 10: 709–720.[Abstract]
4. Suzuki YJ, Forman HJ, Sevanian A. Oxidants as stimulators of signal transduction. Free Radic Biol Med. 1997; 22: 269–285.[CrossRef][Medline] [Order article via Infotrieve]
5. Wolin MS. Reactive oxygen species and vascular signal transduction mechanisms. Microcirculation. 1996; 3: 1–17.[Medline] [Order article via Infotrieve]
6. Griendling KK, Minieri CA, Ollerenshaw JD, Alexander RW. Angiotensin II stimulates NADH and NADPH oxidase activity in cultured vascular smooth muscle cells. Circ Res. 1994; 74: 1141–1148.
7. Irani K, Xia Y, Zweier JL, Sollott SJ, Der CJ, Fearon ER, Sundaresan M, Finkel T, Goldschmidt-Clermont PJ. Mitogenic signaling mediated by oxidants in Ras-transformed fibroblasts. Science. 1997; 275: 1649–1652.
8. Schreck R, Rieber P, Baeuerle PA. Reactive oxygen intermediates as apparently widely used messengers in the activation of the NF-kappa B transcription factor and HIV-1. EMBO J. 1991; 10: 2247–2258.[Medline] [Order article via Infotrieve]
9. Sundaresan M, Yu ZX, Ferrans VJ, Irani K, Finkel T. Requirement for generation of H2O2 for platelet-derived growth factor signal transduction. Science. 1995; 270: 296–299.
10. Suzuki YJ, Ford GD. Superoxide stimulates IP3-induced Ca2+ release from vascular smooth muscle sarcoplasmic reticulum. Am J Physiol. 1992; 262: H114–H116.[Medline] [Order article via Infotrieve]
11. Suzuki YJ, Mizuno M, Packer L. Signal transduction for nuclear factor-kappa B activation. Proposed location of antioxidant-inhibitable step. J Immunol. 1994; 153: 5008–5015.[Abstract]
12. Moran LK, Gutteridge JM, Quinlan GJ. Thiols in cellular redox signalling and control. Curr Med Chem. 2001; 8: 763–772.[Medline] [Order article via Infotrieve]
13. Pessah IN, Feng W. Functional role of hyperreactive sulfhydryl moieties within the ryanodine receptor complex. Antioxid Redox Signal. 2000; 2: 17–25.[Medline] [Order article via Infotrieve]
14. Sen CK. Cellular thiols and redox-regulated signal transduction. Curr Top Cell Regul. 2000; 36: 1–30.[Medline] [Order article via Infotrieve]
15. Lee JW, Helmann JD. The PerR transcription factor senses H2O2 by metal-catalysed histidine oxidation. Nature. 2006; 440: 363–367.[CrossRef][Medline] [Order article via Infotrieve]
16. Yanagisawa M, Kurihara H, Kimura S, Tomobe Y, Kobayashi M, Mitsui Y, Yazaki Y, Goto K, Masaki T. A novel potent vasoconstrictor peptide produced by vascular endothelial cells. Nature. 1988; 332: 411–415.[CrossRef][Medline] [Order article via Infotrieve]
17. Battistini B, Chailler P, D’Orleans-Juste P, Briere N, Sirois P. Growth regulatory properties of endothelins. Peptides. 1993; 14: 385–399.[CrossRef][Medline] [Order article via Infotrieve]
18. Giaid A, Yanagisawa M, Langleben D, Michel RP, Levy R, Shennib H, Kimura S, Masaki T, Duguid WP, Stewart DJ. Expression of endothelin-1 in the lungs of patients with pulmonary hypertension. N Engl J Med. 1993; 328: 1732–1739.
19. Channick RN, Simonneau G, Sitbon O, Robbins IM, Frost A, Tapson VF, Badesch DB, Roux S, Rainisio M, Bodin F, Rubin LJ. Effects of the dual endothelin-receptor antagonist bosentan in patients with pulmonary hypertension: a randomised placebo-controlled study. Lancet. 2001; 358: 1119–1123.[CrossRef][Medline] [Order article via Infotrieve]
20. Rubin LJ, Badesch DB, Barst RJ, Galie N, Black CM, Keogh A, Pulido T, Frost A, Roux S, Leconte I, Landzberg M, Simonneau G. Bosentan therapy for pulmonary arterial hypertension. N Engl J Med. 2002; 346: 896–903.
21. Hassoun PM, Thappa V, Landman MJ, Fanburg BL. Endothelin 1: mitogenic activity on pulmonary artery smooth muscle cells and release from hypoxic endothelial cells. Proc Soc Exp Biol Med. 1992; 199: 165–170.[CrossRef][Medline] [Order article via Infotrieve]
22. Hirata Y, Takagi Y, Fukuda Y, Marumo F. Endothelin is a potent mitogen for rat vascular smooth muscle cells. Atherosclerosis. 1989; 78: 225–228.[CrossRef][Medline] [Order article via Infotrieve]
23. Janakidevi K, Fisher MA, Del Vecchio PJ, Tiruppathi C, Figge J, Malik AB. Endothelin-1 stimulates DNA synthesis and proliferation of pulmonary artery smooth muscle cells. Am J Physiol. 1992; 263: C1295–C1301.[Medline] [Order article via Infotrieve]
24. Davie N, Haleen SJ, Upton PD, Polak JM, Yacoub MH, Morrell NW, Wharton J. ET(A) and ET(B) receptors modulate the proliferation of human pulmonary artery smooth muscle cells. Am J Respir Crit Care Med. 2002; 165: 398–405.
25. Yamboliev IA, Hruby A, Gerthoffer WT. Endothelin-1 activates MAP kinases and c-Jun in pulmonary artery smooth muscle. Pulm Pharmacol Ther. 1998; 11: 205–208.[CrossRef][Medline] [Order article via Infotrieve]
26. Suzuki YJ, Day RM, Tan CC, Sandven TH, Liang Q, Molkentin JD, Fanbury BL. Activation of GATA-4 by serotonin in pulmonary artery smooth muscle cells. J Biol Chem. 2003; 278: 17525–17531.
27. Wedgwood S, Dettman RW, Black SM. ET-1 stimulates pulmonary arterial smooth muscle cell proliferation via induction of reactive oxygen species. Am J Physiol. 2001; 281: L1058–L1067.
28. Lee SL, Wang WW, Moore BJ, Fanburg BL. Dual effect of serotonin on growth of bovine pulmonary artery smooth muscle cells in culture. Circ Res. 1991; 68: 1362–1368.
29. Ihara M, Noguchi K, Saeki T, Fukuroda T, Tsuchida S, Kimura S, Fukami T, Ishikawa K, Nishikibe M, Yano M. Biological profiles of highly potent novel endothelin antagonists selective for the ETA receptor. Life Sci. 1992; 50: 247–255.[CrossRef][Medline] [Order article via Infotrieve]
30. Ishikawa K, Ihara M, Noguchi K, Mase T, Mino N, Saeki T, Fukuroda T, Fukami T, Ozaki S, Nagase T, Nishikibe M, Yano M. Biochemical and pharmacological profile of a potent and selective endothelin B-receptor antagonist, BQ-788. Proc Natl Acad Sci U S A. 1994; 91: 4892–4896.
31. Stadtman ER, Levine RL. Protein oxidation. Ann N Y Acad Sci. 2000; 899: 191–208.[Medline] [Order article via Infotrieve]
32. Rhee SG. Cell signaling. H2O2, a necessary evil for cell signaling. Science. 2006; 312: 1882–1883.
33. Sies H. Ebselen, a selenoorganic compound as glutathione peroxidase mimic. Free Radic Biol Med. 1993; 14: 313–323.[CrossRef][Medline] [Order article via Infotrieve]
34. Levine RL. Proteolysis induced by metal-catalyzed oxidation. Revis Biol Celular. 1989; 21: 347–360.[Medline] [Order article via Infotrieve]
35. Grune T, Reinheckel T, Davies KJ. Degradation of oxidized proteins in mammalian cells. FASEB J. 1997; 11: 526–534.[Abstract]
36. Arner ES, Bjornstedt M, Holmgren A. 1-Chloro-2,4-dinitrobenzene is an irreversible inhibitor of human thioredoxin reductase. Loss of thioredoxin disulfide reductase activity is accompanied by a large increase in NADPH oxidase activity. J Biol Chem. 1995; 270: 3479–3482.
37. Canaider S, Solito E, de Cooupade C, Flower RJ, Russo-Marie F, Goulding NJ, Perretti M. Increased apoptosis in U937 cells over-expressing lipocortin 1 (annexin I). Life Sci. 2000; 66:P L265–PL270.[CrossRef]
38. Solito E, de Coupade C, Canaider S, Goulding NJ, Perretti M. Transfection of annexin 1 in monocytic cells produces a high degree of spontaneous and stimulated apoptosis associated with caspase-3 activation. Br J Pharmacol. 2001; 133: 217–228.[CrossRef][Medline] [Order article via Infotrieve]
39. Debret R, El Btaouri H, Duca L, Rahman I, Radke S, Haye B, Sallenave JM, Antonicelli F. Annexin A1 processing is associated with caspase-dependent apoptosis in BZR cells. FEBS Lett. 2003; 546: 195–202.[CrossRef][Medline] [Order article via Infotrieve]
40. Alldridge LC, Bryant CE. Annexin 1 regulates cell proliferation by disruption of cell morphology and inhibition of cyclin D1 expression through sustained activation of the ERK1/2 MAPK signal. Exp Cell Res. 2003; 290: 93–107.[CrossRef][Medline] [Order article via Infotrieve]
41. Kim SB, Yang WS, Lee OS, Lee KP, Park JS, Na DS. Lipocortin-1 inhibits proliferation of cultured human mesangial cells. Nephron. 1996; 74: 39–44.[Medline] [Order article via Infotrieve]
42. Roederer M, Staal FJ, Raju PA, Ela SW, Herzenberg LA, Herzenberg LA. Cytokine-stimulated human immunodeficiency virus replication is inhibited by N-acetyl-L-cysteine. Proc Natl Acad Sci U S A. 1990; 87: 4884–4888.
43. Staal FJ, Roederer M, Herzenberg LA, Herzenberg LA. Intracellular thiols regulate activation of nuclear factor kappa B and transcription of human immunodeficiency virus. Proc Natl Acad Sci U S A. 1990; 87: 9943–9947.
44. Rao GN, Berk BC. Active oxygen species stimulate vascular smooth muscle cell growth and proto-oncogene expression. Circ Res. 1992; 70: 593–599.
45. Lee SL, Wang WW, Fanburg BL. Superoxide as an intermediate signal for serotonin-induced mitogenesis. Free Rad Biol Med. 1998; 24: 855–858.[CrossRef][Medline] [Order article via Infotrieve]
46. Lee SL, Wang WW, Finlay GA, Fanburg BL. Serotonin stimulates mitogen-activated protein kinase activity through the formation of superoxide anion. Am J Physiol. 1999; 277: L282–L291.[Medline] [Order article via Infotrieve]
47. Lee SL, Simon AR, Wang WW, Fanburg BL. H2O2 signals 5-HT-induced ERK MAP kinase activation and mitogenesis of smooth muscle cells. Am J Physiol. 2001; 281: L646–L652.
48. Lawrie A, Spiekerkoetter E, Martinez EC, Ambartsumian N, Sheward WJ, MacLean MR, Harmar AJ, Schmidt AM, Lukanidin E, Rabinovitch M. Interdependent serotonin transporter and receptor pathways regulate S100A4/Mts1, a gene associated with pulmonary vascular disease. Circ Res. 2005; 97: 227–235.
49. Ushio-Fukai M, Zafari AM, Fukui T, Ishizaka N, Griendling KK. p22phox is a critical component of the superoxide-generating NADH/NADPH oxidase system and regulates angiotensin II-induced hypertrophy in vascular smooth muscle cells. J Biol Chem. 1996; 271: 23317–23321.
50. Kondo N, Nakamura H, Masutani H, Yodoi J. Redox regulation of human thioredoxin network. Antioxid Redox Signal. 2006; 8: 1881–1890.[CrossRef][Medline] [Order article via Infotrieve]
51. Dalle-Donne I, Aldini G, Carini M, Colombo R, Rossi R, Milzani A. Protein carbonylation, cellular dysfunction, and disease progression. J Cell Mol Med. 2006; 10: 389–406.[Medline] [Order article via Infotrieve]
52. Suzuki YJ, Nagase H, Wong CM, Vinod Kumar S, Jain V, Park AM, Day RM. Regulation of Bcl-xL expression in lung vascular smooth muscle. Am J Respir Cell Mol Biol. 2007; 36: 678–687.
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