Detection ofS-Nitrosothiols and Other Nitric Oxide Derivatives by Photolysis–Chemiluminescence Spectrometry

Alpert, Caroline S.; Ramdev, Nomith; George, D; Loscalzo, Joseph

doi:10.1006/abio.1996.9947

Cited by 32 publications

(28 citation statements)

References 11 publications

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“…A variety of indirect methods have been used to identify S-nitrosothiols and S-nitrosoproteins, including S-nitroso-serum albumin (1), Snitroso-hemoglobin (18), S-nitroso-ryanodine receptor (19), and S-nitroso-caspase-3 (20). Photolysis-chemiluminescence (1,21) has been used by our group and others, and, although sensitive, has limited specificity because of interference by nitrite (22). In addition, this and other indirect chemical methods suffer from post hoc formation of S-nitrosothiols, occurring during any step at which the pH is lowered below 7.4 in the presence of nitrite.…”

Section: Discussionmentioning

confidence: 99%

S -nitrosoprotein formation and localization in endothelial cells

Yang

Loscalzo

2004

Proc. Natl. Acad. Sci. U.S.A.

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131

109

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Protein S-nitrosation represents a recently described form of post-translational modification that is rapid and reversible. However, the analysis of protein S-nitrosation in situ has been difficult because of the absence of specific probes and the instability of cellular protein S -nitrosothiols. We developed a rapid and specific method for detecting endothelial S -nitrosoproteins patterned after the biotin switch method that involves thiol alkylation followed by reductive generation of thiols from S -nitrosothiols, which are then labeled with either a biotin- or Texas red-derivative of methanethiosulfonate. When we used this methodology, we found that S-nitrosated proteins can form within endothelial cells from an exogenous S -nitrosothiol donor or from endogenous production of NO by endothelial NO synthase. When we used confocal microscopy, we found that these S -nitrosoproteins exist mainly in the mitochondria and peri-mitochondrial compartment, and that their half-life is ≈1 h. Cellular S-nitrosated protein abundance changed as expected, with changes in activity of NO synthase, and with impairment of mitochondrial function and scavenging of peroxynitrite. We used a proteomic approach involving two-dimensional gel electrophoresis and mass spectrometry, and found that a limited number of S -nitrosoproteins exist in endothelial cells ( S -nitrosoproteome) and identified GAPDH, vimentin, β-galactosidase, peroxiredoxin 1, β-actin, and ubiquitin-conjugating enzyme E2 among them. The most abundant S-nitrosated protein in the resting endothelial cell is GAPDH, suggesting a regulatory function for NO in glycolysis. These data offer methods and insights into identifying the protein targets of S-nitrosation reactions and their potential role in cell function and phenotype.

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Section: Discussionmentioning

confidence: 99%

S -nitrosoprotein formation and localization in endothelial cells

Yang

Loscalzo

2004

Proc. Natl. Acad. Sci. U.S.A.

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131

109

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“…Free · NO and · NO-adducts, including SNOs, N-nitroso compounds (RNNOs), and metalnitrosyls can contribute to signal output of photolysis with chemiluminescent detection [26]. Under conditions found in the A. stephensi midgut, it is likely that free · NO reacts rapidly to generate a variety of · NO metabolites.…”

Section: Discussionmentioning

confidence: 99%

“…Photolysis coupled with chemiluminescent detection is a sensitive measure of SNOs and metalnitrosyl adducts and to some extent N-nitroso compounds [19]. Photolyzed-· NO levels in midgut lysates were measured immediately after (0h) to 33.5h after A. stephensi feeding on uninfected or P. berghei-infected mice.…”

Section: Photolyzable Adducts Of · No In the Mosquito Midgut Are Incrmentioning

confidence: 99%

Nitric oxide metabolites induced in Anopheles stephensi control malaria parasite infection

Peterson

Gow

Luckhart

2007

Free Radical Biology and Medicine

100

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Malaria parasite infection in anopheline mosquitoes is limited by inflammatory levels of nitric oxide metabolites. To assess the mechanisms of parasite stasis or toxicity, we investigated the biochemistry of these metabolites within the blood-filled mosquito midgut. Our data indicate that nitrates, but not nitrites, are elevated in the Plasmodium-infected midgut. Although levels of S-nitrosothiols do not change with infection, blood proteins are S-nitrosylated after ingestion by the mosquito. In addition, photolyzable nitric oxide, which can be attributed to metal nitrosyls, is elevated following infection and, based on the abundance of hemoglobin, likely includes heme iron nitrosyl. The persistance of oxyhemoglobin throughout blood digestion and changes in hemoglobin conformation in response to infection suggest that hemoglobin catalyzes the synthesis of nitric oxide metabolites in a reducing environment. Provision of urate, a potent reductant and scavenger of oxidants and nitrating agents, as a dietary supplement to mosquitoes increased parasite infection levels relative to allantoin-fed controls, suggesting that nitrosative and/or oxidative stresses negatively impact developing parasites. Collectively, our results reveal a unique role for nitric oxide in an oxyhemoglobin-rich environment. In contrast to facilitating oxygen delivery by hemoglobin in the mammalian vasculature, nitric oxide synthesis in the blood-filled mosquito midgut drives the formation of toxic metabolites that limit parasite development.

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“…The method's linearity for various S-nitrosothiols including S-nitroso-bovine serum albumin, which was prepared from bovine serum albumin with acidified nitrite (Stamler et al, 1992b), sensitivity as well as intra-assay variability were reported, but data on the method's accuracy were missing (Stamler et al, 1992a). The photolysis-chemiluminescence method originally used to quantitate S-nitrosothiols in human plasma (Stamler et al, 1992a) was reported several years later in detail by the same group (Alpert et al, 1997). Unfortunately, in that study no S-nitrosothiol levels was determined in human plasma.…”

Section: S -Nitrosothiolsmentioning

confidence: 97%

Quantitative Determination of Free and Protein‐Associated 3‐Nitrotyrosine and S ‐Nitrosothiols in the Circulation by Mass Spectrometry and Other Methodologies: A Critical Review and Discussion from the Analytical and Review Point of View

Tsikas¹

2006

Redox Proteomics

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Detection ofS-Nitrosothiols and Other Nitric Oxide Derivatives by Photolysis–Chemiluminescence Spectrometry

Cited by 32 publications

References 11 publications

S -nitrosoprotein formation and localization in endothelial cells

S -nitrosoprotein formation and localization in endothelial cells

Nitric oxide metabolites induced in Anopheles stephensi control malaria parasite infection

Quantitative Determination of Free and Protein‐Associated 3‐Nitrotyrosine and S ‐Nitrosothiols in the Circulation by Mass Spectrometry and Other Methodologies: A Critical Review and Discussion from the Analytical and Review Point of View

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