Trial reportThe Journal of biological chemistry2014
Assessment of myeloperoxidase activity by the conversion of hydroethidine to 2-chloroethidium.
Trial report in The Journal of biological chemistry, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
22 citing papers in PubMed.
- Challenges in myeloperoxidase quantification: the inadequacies of current colorimetric approaches.Frontiers in molecular biosciences · 2025Article
- Molecular magnetic resonance imaging of myeloperoxidase activity identifies culprit lesions and predicts future atherothrombosis.European heart journal. Imaging methods and practice · 2024Article
- Intraplaque Myeloperoxidase Activity as Biomarker of Unstable Atheroma and Adverse Clinical Outcomes in Human Atherosclerosis.JACC. Advances · 2023Article
- Mechanisms of Oxidative Stress and Therapeutic Targets following Intracerebral Hemorrhage.Oxidative medicine and cellular longevity · 2021Review
- Myeloperoxidase: a potential therapeutic target for coronary artery disease.Expert opinion on therapeutic targets · 2020Review
- Tracking isotopically labeled oxidants using boronate-based redox probes.The Journal of biological chemistry · 2020Article
- Boronate-Based Probes for Biological Oxidants: A Novel Class of Molecular Tools for Redox Biology.Frontiers in chemistry · 2020Review
- Pharmacological characterization of the seven human NOX isoforms and their inhibitors.Redox biology · 2019Article
- Article
- Small-molecule luminescent probes for the detection of cellular oxidizing and nitrating species.Free radical biology & medicine · 2018Review
- Monitoring the Instant Creation of a New Fluorescent Signal for Evaluation of DNA Conformation Based on Intercalation Complex.Journal of fluorescence · 2018Article
- Targeting myeloperoxidase in inflammatory atherosclerosis.European heart journal · 2018Article
- Detection and Characterization of Reactive Oxygen and Nitrogen Species in Biological Systems by Monitoring Species-Specific Products.Antioxidants & redox signaling · 2018Review
- Recent Developments in the Probes and Assays for Measurement of the Activity of NADPH Oxidases.Cell biochemistry and biophysics · 2017Article
- Regulation of Cellular Redox Signaling by Matricellular Proteins in Vascular Biology, Immunology, and Cancer.Antioxidants & redox signaling · 2017Article
- Recent developments in detection of superoxide radical anion and hydrogen peroxide: Opportunities, challenges, and implications in redox signaling.Archives of biochemistry and biophysics · 2017Review
- The roles of myeloperoxidase in coronary artery disease and its potential implication in plaque rupture.Redox report : communications in free radical research · 2017Review
- A Critical Review of Methodologies to Detect Reactive Oxygen and Nitrogen Species Stimulated by NADPH Oxidase Enzymes: Implications in Pesticide Toxicity.Current pharmacology reports · 2016Article
- Fast and Specific Assessment of the Halogenating Peroxidase Activity in Leukocyte-enriched Blood Samples.Journal of visualized experiments : JoVE · 2016Article
- Imaging Reactive Oxygen Species-Induced Modifications in Living Systems.Antioxidants & redox signaling · 2016Review
Corrections and comments
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Authors and funding
8 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Oxidants derived from myeloperoxidase (MPO) contribute to inflammatory diseases. In vivo MPO activity is commonly assessed by the accumulation of 3-chlorotyrosine (3-Cl-Tyr), although 3-Cl-Tyr is formed at low yield and is subject to metabolism. Here we show that MPO activity can be assessed using hydroethidine (HE), a probe commonly employed for the detection of superoxide. Using LC/MS/MS, (1)H NMR, and two-dimensional NOESY, we identified 2-chloroethidium (2-Cl-E(+)) as a specific product when HE was exposed to hypochlorous acid (HOCl), chloramines, MPO/H2O2/chloride, and activated human neutrophils. The rate constant for HOCl-mediated conversion of HE to 2-Cl-E(+) was estimated to be 1.5 × 10(5) M(-1)s(-1). To investigate the utility of 2-Cl-E(+) to assess MPO activity in vivo, HE was injected into wild-type and MPO-deficient (Mpo(-/-)) mice with established peritonitis or localized arterial inflammation, and tissue levels of 2-Cl-E(+) and 3-Cl-Tyr were then determined by LC/MS/MS. In wild-type mice, 2-Cl-E(+) and 3-Cl-Tyr were detected readily in the peritonitis model, whereas in the arterial inflammation model 2-Cl-E(+) was present at comparatively lower concentrations (17 versus 0.3 pmol/mg of protein), and 3-Cl-Tyr could not be detected. Similar to the situation with 3-Cl-Tyr, tissue levels of 2-Cl-E(+) were decreased substantially in Mpo(-/-) mice, indicative of the specificity of the assay. In the arterial inflammation model, 2-Cl-E(+) was absent from non-inflamed arteries and blood, suggesting that HE oxidation occurred locally in the inflamed artery. Our data suggest that the conversion of exogenous HE to 2-Cl-E(+) may be a useful selective and sensitive marker for MPO activity in addition to 3-Cl-Tyr.
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