Detection of Nitric Oxide by Electron Paramagnetic Resonance Spectroscopy: Spin-Trapping with Iron-Dithiocarbamates.
Electron paramagnetic resonance (EPR) spectroscopy is the ideal methodology to identify radicals (detection and characterization of molecular structure) and to study their kinetics, in both simple and complex biological systems. The very low concentration and short life-time of NO and of many other radicals do not favor its direct detection and spin-traps are needed to produce a new and persistent radical that can be subsequently detected by EPR spectroscopy.In this chapter, we present the basic concepts of EPR spectroscopy and of some spin-trapping methodologies to study NO. The "strengths and weaknesses" of iron-dithiocarbamates utilization, the NO traps of choice for the authors, are thoroughly discussed and a detailed description of the method to quantify the NO formation by molybdoenzymes is provided.
UCIBIO, REQUIMTE, Departamento Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Campus de Caparica, 2829-516, Caparica, Portugal. luisa.maia@fct.unl.pt.
Luisa B
LB
Maia
UCIBIO, REQUIMTE, Departamento Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Campus de Caparica, 2829-516, Caparica, Portugal.
José J G
JJ
Moura
D000818:Animals; D004578:Electron Spin Resonance Spectroscopy; D006801:Humans; D007501:Iron / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism*; D013113:Spin Labels; D013859:Thiocarbamates / Q000378:metabolism*
D013113:Spin Labels; D013859:Thiocarbamates; D009569:Nitric Oxide; D007501:Iron
Aldehyde oxidoreductase; Electron paramagnetic resonance (EPR); Iron-dithiocarbamate; Nitric oxide radical; Nitrite; Spin-trap; Xanthine oxidoreductase