Chemical Interaction of Nitric Oxide With Protein Thiols

Nitric oxide (NO) is produced by different isoforms of NO-synthases and operates as a mediator of important cell signaling pathways. To explore thiol-based protein modifications in a situation of nitrosative stress, two transgenic mouse models recently generated in our laboratory were used: Cardiac specific overexpression of inducible nitric oxide synthase (iNOS) (tg-iNOS + ) and tg-iNOS + with concomitant myoglobin-deficiency (tg-iNOS + /myo −/− ). Protein S-nitrosation, an important redox-based posttranslational modification, revealed no differences between WT and tg-iNOS + hearts as measured by the biotin-switch assay and 2-D PAGE. Even in the absence of myoglobin - an efficient endogenous NO-oxidase - the protein S-nitrosation pattern for nearly all the detected proteins (>40) remained unchanged in the tg-iNOS + /myo −/− hearts, with the exception of three proteins. Tandem mass spectrometry uncovered these proteins as peroxiredoxins (Prx II, III, VI), which are known to possess peroxidase activity, whereby hydrogen peroxide, peroxynitrite and a wide range of organic hydroperoxides are reduced and detoxified. To prove whether the higher abundance of the Prxs was due to enhanced S-nitrosation or due to changes in their basal expression levels, immunoblotting with specific antibodies was applied and revealed upregulation of Prx VI in tg-iNOS + /myo −/− hearts. The other proteins found to be S-nitrosated were identified as well. Data mining indicated a significant overlap of these proteins with proteins becoming glutathiolated. Protein glutathiolation detected by immunoblotting was enhanced in the tg-iNOS + hearts and even more so in the tg-iNOS + /myo −/− hearts. We conclude that protein glutathiolation in our transgenic model of nitrosative stress is important to protect protein thiols from irreversible oxidation. The upregulation of antioxidant proteins like Prx VI appears to be an additional mechanism to antagonize an excess of reactive oxygen/nitrogen species. Enhanced S-nitrosation of the Prxs may serve a new function in the signalling cascade coping with nitrosative stress.

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Nitric Oxide Alleviates Iron Toxicity by Reducing Oxidative Damage and Growth Inhibition in Wheat (Triticum aestivum L.) Seedlings

International Journal of Plant and Environment ◽

10.18811/ijpen.v5i01.3 ◽

2019 ◽

Vol 5 (01) ◽

pp. 16-22

Author(s):

Nalini Pandey ◽

Laxmi Verma

Keyword(s):

Oxidative Stress ◽

Nitric Oxide ◽

Oxidative Damage ◽

Sodium Nitroprusside ◽

Dry Weight ◽

Triticum Aestivum L ◽

Tolerance Index ◽

Promoting Effect ◽

Wheat Seedlings ◽

Protein Thiols

Nitric oxide (NO) is an important bioactive signaling molecule in plants which modulates a variety of physiological processes and responses to abiotic and biotic stresses. In this study, the effects of exogenous NO supplied as sodium nitroprusside (SNP) in wheat seedlings under ironinduced oxidative damage was investigated. An appropriate concentration of NO was determined by conducting a preliminary experiment. In solution culture, wheat seeds were grown in the control (100 μM Fe), and toxic Fe (400 μM Fe) levels and the toxic Fe supply was treated with various levels of (50, 100, 200 and 500 μM) sodium nitroprusside (SNP). The results indicated that 400 μM Fe significantly decreased percentage germination, tolerance index, root lengths as well as fresh and dry weight compared to control. Exogenous SNP attenuated the inhibition of wheat seed germination. The promoting effect was most pronounced at 100 μM SNP. The accumulated concentration of iron and active Fe was significantly decreased by SNP treated Fe toxic seedlings. Toxicity of Fe caused oxidative stress by elevating hydrogen peroxide (H2O2), malondialdehyde (MDA) and proline contents in roots of wheat seedlings. One hundred μM SNP counteracted Fe toxicity by reducing the H2O2, MDA and proline contents of toxic Fe exposed seedlings. Meanwhile, application of SNP markedly reduced the activities of superoxide dismutases (SOD), catalases (CAT), peroxidase (POD), ascorbate peroxidases (APX), non protein thiols (NPT) and of glutathione reductase (GR) and increased ascorbate (ASc) compared with Fe toxic treatment alone, thereby indicating the modulation of the antioxidative capacity in the root under Fe stress by NO. The results indicated that the exogenous application of SNP, improved the antioxidant enzymes activity of wheat seedlings against Fe induced oxidative stress.

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Role of Ascorbate and Protein Thiols in the Release of Nitric oxide from S-Nitroso-Albumin and S-Nitroso-Glutathione in Human Plasma

Free Radical Biology and Medicine ◽

10.1016/s0891-5849(96)00378-4 ◽

1997 ◽

Vol 22 (4) ◽

pp. 633-642 ◽

Cited By ~ 89

Author(s):

Giuseppe Scorza ◽

Donatella Pietraforte ◽

Maurizio Minetti

Keyword(s):

Nitric Oxide ◽

Human Plasma ◽

Protein Thiols

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Glutathione and Nitric Oxide: Key Team Players in Use and Disuse of Skeletal Muscle

Nutrients ◽

10.3390/nu11102318 ◽

2019 ◽

Vol 11 (10) ◽

pp. 2318 ◽

Cited By ~ 4

Author(s):

Sara Baldelli ◽

Fabio Ciccarone ◽

Dolores Limongi ◽

Paola Checconi ◽

Anna Teresa Palamara ◽

...

Keyword(s):

Nitric Oxide ◽

Skeletal Muscle ◽

Contractile Activity ◽

Direct Consequence ◽

Redox Status ◽

No Production ◽

Skeletal Muscle Function ◽

Protein Thiols ◽

Characteristic Features ◽

Contraction Regulation

Glutathione (GSH) is the main non-enzymatic antioxidant playing an important role in detoxification, signal transduction by modulation of protein thiols redox status and direct scavenging of radicals. The latter function is not only performed against reactive oxygen species (ROS) but GSH also has a fundamental role in buffering nitric oxide (NO), a physiologically-produced molecule having-multifaceted functions. The efficient rate of GSH synthesis and high levels of GSH-dependent enzymes are characteristic features of healthy skeletal muscle where, besides the canonical functions, it is also involved in muscle contraction regulation. Moreover, NO production in skeletal muscle is a direct consequence of contractile activity and influences several metabolic myocyte pathways under both physiological and pathological conditions. In this review, we will consider the homeostasis and intersection of GSH with NO and then we will restrict the discussion on their role in processes related to skeletal muscle function and degeneration.

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Continuous exposure to high concentrations of nitric oxide leads to persistent inhibition of oxygen consumption by J774 cells as well as extraction of oxygen by the extracellular medium

Biochemical Journal ◽

10.1042/bj3460407 ◽

2000 ◽

Vol 346 (2) ◽

pp. 407-412 ◽

Cited By ~ 20

Author(s):

Antonia ORSI ◽

Belén BELTRÁN ◽

Emilio CLEMENTI ◽

Katarina HALLÉN ◽

Martin FEELISCH ◽

...

Keyword(s):

Nitric Oxide ◽

Oxygen Consumption ◽

Chemical Interaction ◽

Continuous Exposure ◽

Cell Respiration ◽

Extracellular Medium ◽

Pathological Conditions ◽

J774 Cells ◽

High Concentrations ◽

Complex I Activity

Nitric oxide (NO) plays a key role in many physiological and pathophysiological events, including the control of cell respiration. Both reversible and irreversible inhibition of mitochondrial respiration have been reported following the generation of NO by cells. We have exposed the murine macrophage cell line J774 to high concentrations of NO, such as are generated in some pathological conditions, and determined their effect on oxygen consumption. We observed a persistent inhibition of respiration which was due to a redox-dependent, progressive inhibition of complex I activity. No other enzyme of the respiratory chain was inhibited in this way. At the same time, we detected a paradoxical removal of oxygen by the extracellular medium. This removal was due to a chemical interaction between dissolved oxygen and NO-related species released from cells exposed to NO. A similar removal of oxygen by the cell supernatant also occurred following activation of cells with cytokines and bacterial products. Thus, the amounts of NO generated during pathological conditions may contribute to tissue hypoxia both by inhibiting cell respiration and by promoting removal of oxygen from the extracellular medium.

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Interaction of Hydrogen Sulfide with Nitric Oxide in the Cardiovascular System

Oxidative Medicine and Cellular Longevity ◽

10.1155/2016/6904327 ◽

2016 ◽

Vol 2016 ◽

pp. 1-16 ◽

Cited By ~ 71

Author(s):

B. V. Nagpure ◽

Jin-Song Bian

Keyword(s):

Nitric Oxide ◽

Hydrogen Sulfide ◽

Cardiovascular System ◽

Chemical Interaction ◽

New Family ◽

Pathophysiological Role ◽

Effective Role ◽

No Signaling ◽

The Individual ◽

Heart Contractility

Historically acknowledged as toxic gases, hydrogen sulfide (H2S) and nitric oxide (NO) are now recognized as the predominant members of a new family of signaling molecules, “gasotransmitters” in mammals. While H2S is biosynthesized by three constitutively expressed enzymes (CBS, CSE, and 3-MST) from L-cysteine and homocysteine, NO is generated endogenously from L-arginine by the action of various isoforms of NOS. Both gases have been transpired as the key and independent regulators of many physiological functions in mammalian cardiovascular, nervous, gastrointestinal, respiratory, and immune systems. The analogy between these two gasotransmitters is evident not only from their paracrine mode of signaling, but also from the identical and/or shared signaling transduction pathways. With the plethora of research in the pathophysiological role of gasotransmitters in various systems, the existence of interplay between these gases is being widely accepted. Chemical interaction between NO and H2S may generate nitroxyl (HNO), which plays a specific effective role within the cardiovascular system. In this review article, we have attempted to provide current understanding of the individual and interactive roles of H2S and NO signaling in mammalian cardiovascular system, focusing particularly on heart contractility, cardioprotection, vascular tone, angiogenesis, and oxidative stress.

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Electrospray mass-spectrometric, spectrophotometric and electrochemical methods do not provide evidence for the binding of nitric oxide by pyocyanine at pH7

Biochemical Journal ◽

10.1042/bj3220025 ◽

1997 ◽

Vol 322 (1) ◽

pp. 25-29 ◽

Cited By ~ 15

Author(s):

Dragic V. VUKOMANOVIC ◽

Dick E. ZOUTMAN ◽

John A. STONE ◽

Gerald S. MARKS ◽

James F. BRIEN ◽

...

Keyword(s):

Nitric Oxide ◽

Mechanism Of Action ◽

Chemical Interaction ◽

Mass Spectrometric ◽

Electrochemical Methods ◽

Neutral Ph

In several recent publications on pyocyanine, its mechanism of action has been attributed to an ability to react with nitric oxide (NO), resulting in the formation of an adduct. We examined the chemical interaction of pyocyanine and NO using electrospray (ES) MS, spectrophotometry and voltammetry at neutral pH and with 10Ő100 ƁM pyocyanine. No binding of NO to pyocyanine was observed. Alternative mechanisms for the inhibition of NO-induced vasorelaxation by pyocyanine should be sought.

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Direct and Indirect Inhibition of Salmonella Peptide Deformylase by Nitric Oxide

mBio ◽

10.1128/mbio.01383-20 ◽

2020 ◽

Vol 11 (6) ◽

Author(s):

Anshika Singhal ◽

Ferric C. Fang

Keyword(s):

Nitric Oxide ◽

Metal Binding ◽

Nitrosative Stress ◽

Peptide Deformylase ◽

Zinc Toxicity ◽

No Production ◽

Bacterial Cells ◽

Intracellular Zinc ◽

Content Type ◽

Protein Thiols

ABSTRACT Salmonella enterica serovar Typhimurium is an intracellular pathogen that elicits nitric oxide (NO·) production by host macrophages. NO· is a potent antimicrobial mediator with diverse targets, including protein thiols and metal centers. The mobilization of zinc from metalloproteins by NO· increases the availability of free intracellular zinc, which is detrimental to bacterial cells, but the precise mechanism of zinc cytotoxicity is uncertain. Here, we show that excess zinc results in the mismetallation of the essential iron-containing enzyme peptide deformylase (PDF), thereby diminishing its activity. PDF mismetallation is observed in zinc-treated bacteria lacking the zinc exporters ZntA and ZitB and is also observed during nitrosative stress, suggesting that NO·-mediated zinc mobilization results in PDF mismetallation. However, NO· also inhibits PDF directly by S-nitrosylating the metal-binding Cys90 residue. These observations identify PDF as an essential bacterial protein that is subject to both direct and indirect inactivation by NO·, providing a novel mechanism of zinc toxicity and NO·-mediated antibacterial activity. IMPORTANCE We have previously shown that the host-derived antimicrobial mediator nitric oxide (NO·) mobilizes zinc from bacterial metalloproteins. The present study demonstrates that NO· inactivates the essential iron-containing enzyme peptide deformylase, both by promoting its mismetallation by zinc and by directly modifying its metal-binding site. We explain how free intracellular zinc is detrimental for cells and reveal a new mechanism of NO·-mediated bacterial growth inhibition that is distinct from previously known targets.

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Chemical Interactions at the Interface of Plant Root Hair Cells and Intracellular Bacteria

Microorganisms ◽

10.3390/microorganisms9051041 ◽

2021 ◽

Vol 9 (5) ◽

pp. 1041

Author(s):

Xiaoqian Chang ◽

Kathryn L. Kingsley ◽

James F. White

Keyword(s):

Nitric Oxide ◽

Carbon Dioxide ◽

Chemical Interaction ◽

Plant Root ◽

Plant Cells ◽

Intracellular Bacteria ◽

Chemical Interactions ◽

Abiotic And Biotic Stresses ◽

Biotic Stresses ◽

Root Hair Cells

In this research, we conducted histochemical, inhibitor and other experiments to evaluate the chemical interactions between intracellular bacteria and plant cells. As a result of these experiments, we hypothesize two chemical interactions between bacteria and plant cells. The first chemical interaction between endophyte and plant is initiated by microbe-produced ethylene that triggers plant cells to grow, release nutrients and produce superoxide. The superoxide combines with ethylene to form products hydrogen peroxide and carbon dioxide. In the second interaction between microbe and plant the microbe responds to plant-produced superoxide by secretion of nitric oxide to neutralize superoxide. Nitric oxide and superoxide combine to form peroxynitrite that is catalyzed by carbon dioxide to form nitrate. The two chemical interactions underlie hypothesized nutrient exchanges in which plant cells provide intracellular bacteria with fixed carbon, and bacteria provide plant cells with fixed nitrogen. As a consequence of these two interactions between endophytes and plants, plants grow and acquire nutrients from endophytes, and plants acquire enhanced oxidative stress tolerance, becoming more tolerant to abiotic and biotic stresses.

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