Maia Luisa B, Moura José J G
UCIBIO, REQUIMTE, Departamento Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Campus de Caparica, 2829-516, Caparica, Portugal.
Methods Mol Biol. 2016;1424:81-102. doi: 10.1007/978-1-4939-3600-7_8.
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.
电子顺磁共振（EPR）光谱法是在简单和复杂生物系统中识别自由基（检测和表征分子结构）并研究其动力学的理想方法。一氧化氮（NO）和许多其他自由基的浓度极低且寿命很短，不利于直接检测，因此需要使用自旋捕集剂来产生一种新的持久性自由基，随后可通过EPR光谱法进行检测。在本章中，我们介绍了EPR光谱法的基本概念以及一些用于研究NO的自旋捕集方法。我们深入讨论了二硫代氨基甲酸盐铁的“优缺点”，这是作者选择的NO捕集剂，并详细描述了通过钼酶定量NO生成的方法。
