Isotope tracing enhancement of chemiluminescence assays for nitric oxide research
-
Julia Cornelius
Abstract
Chemiluminescence assays are used widely for the detection of nitric oxide (NO)-derived species in biological fluids and tissues. Here, we demonstrate that these assays can be interfaced with mass-sensitive detectors for parallel determination of isotopic abundance. Results obtained with tri-iodide and ascorbic acid-based reductive assays indicate that mass spectrometric detection enables NO isotope-tracing experiments to be carried out to a limit of detectability of a few picomoles, a sensitivity similar to that of standard gas phase chemiluminescence methods. The advantage afforded by mass spectrometric detection is demonstrated using the murine macrophage cell line J774, which is shown here to reduce 15NO3- to 15NO2- under anoxic conditions. The particular combination of an analytical and cellular system described here may hold promise for future characterization of the enzymatic pathways contributing to mammalian nitrate reductase activity, without background interference from 14NO2- derived from other sources.
©2009 by Walter de Gruyter Berlin New York
Articles in the same Issue
- Minireview
- Functional genetic mouse models: promising tools for investigation of the proteolytic internet
- Protein Structure and Function
- Primary sequence, together with other factors, influence peptide deimination by peptidylarginine deiminase-4
- Mercury and cadmium trigger expression of the copper importer Ctr1B, which enables Drosophila to thrive on heavy metal-loaded food
- Cell Biology and Signaling
- Interferon-γ-mediated pathways and in vitro PBMC proliferation in HIV-infected patients
- Exploring the pathogenesis of renal cell carcinoma: pathway and bioinformatics analysis of dysregulated genes and proteins
- Aptamers selected against the unglycosylated EGFRvIII ectodomain and delivered intracellularly reduce membrane-bound EGFRvIII and induce apoptosis
- Involvement of heparan sulfate proteoglycans in cellular uptake of high molecular weight kininogen
- Overexpression of Pygopus2 protects HeLa cells from vinblastine-induced apoptosis
- Proteolysis
- Selectivity of propeptide-enzyme interaction in cathepsin L-like cysteine proteases
- Murine and human cathepsin B exhibit similar properties: possible implications for drug discovery
- Novel Techniques
- Isotope tracing enhancement of chemiluminescence assays for nitric oxide research
Articles in the same Issue
- Minireview
- Functional genetic mouse models: promising tools for investigation of the proteolytic internet
- Protein Structure and Function
- Primary sequence, together with other factors, influence peptide deimination by peptidylarginine deiminase-4
- Mercury and cadmium trigger expression of the copper importer Ctr1B, which enables Drosophila to thrive on heavy metal-loaded food
- Cell Biology and Signaling
- Interferon-γ-mediated pathways and in vitro PBMC proliferation in HIV-infected patients
- Exploring the pathogenesis of renal cell carcinoma: pathway and bioinformatics analysis of dysregulated genes and proteins
- Aptamers selected against the unglycosylated EGFRvIII ectodomain and delivered intracellularly reduce membrane-bound EGFRvIII and induce apoptosis
- Involvement of heparan sulfate proteoglycans in cellular uptake of high molecular weight kininogen
- Overexpression of Pygopus2 protects HeLa cells from vinblastine-induced apoptosis
- Proteolysis
- Selectivity of propeptide-enzyme interaction in cathepsin L-like cysteine proteases
- Murine and human cathepsin B exhibit similar properties: possible implications for drug discovery
- Novel Techniques
- Isotope tracing enhancement of chemiluminescence assays for nitric oxide research
