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Isotope tracing enhancement of chemiluminescence assays for nitric oxide research
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Cornelius, Julia, Tran, Tuan, Turner, Nicole, Piazza, Abigail, Mills, Lauren, Slack, Ryan, Hauser, Sean, Alexander, J. Steven, Grisham, Matthew B., Feelisch, Martin and Rodriguez, Juan. (2009) Isotope tracing enhancement of chemiluminescence assays for nitric oxide research. Biological Chemistry, Vol.390 (No.2). pp. 181-189. ISSN 1431-6730
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Official URL: http://dx.doi.org/10.1515/BC.2009.017
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 (NO3-)-N-15 to (NO2-)-N-15 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 (NO2-)-N-14 derived from other sources.
| Item Type: | Journal Article |
|---|---|
| Subjects: | Q Science > QD Chemistry |
| Divisions: | Faculty of Medicine > Warwick Medical School > Clinical Sciences Research Institute (CSRI) Faculty of Medicine > Warwick Medical School > Metabolic and Vascular Health Faculty of Medicine > Warwick Medical School |
| Journal or Publication Title: | Biological Chemistry |
| Publisher: | Walter de Gruyter & Co |
| ISSN: | 1431-6730 |
| Date: | February 2009 |
| Volume: | Vol.390 |
| Number: | No.2 |
| Number of Pages: | 9 |
| Page Range: | pp. 181-189 |
| Identification Number: | 10.1515/BC.2009.017 |
| Status: | Peer Reviewed |
| Publication Status: | Published |
| Access rights to Published version: | Restricted or Subscription Access |
| Funder: | Louisiana Biomedical Research Network, National Science Foundation |
| Grant number: | 2 P20 RR016456, 0641516 |
| URI: | http://wrap.warwick.ac.uk/id/eprint/28437 |
Data sourced from Thomson Reuters' Web of Knowledge
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