Zielonka Jacek, Sikora Adam, Adamus Jan, Kalyanaraman B
Department of Biophysics and Free Radical Research Center, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI, 53226, USA,
Methods Mol Biol. 2015;1264:171-81. doi: 10.1007/978-1-4939-2257-4_16.
The development of boronic probes enabled reliable detection and quantitative analysis of hydrogen peroxide and peroxynitrite. The major product, in which boronate moiety of the probe is replaced by the hydroxyl group, is however common for both oxidants. Here, we describe how ortho-isomer of mitochondria-targeted phenylboronic acid can be used to detect and differentiate peroxynitrite-dependent and peroxynitrite-independent probe oxidation. This method highlights the detection and quantification of both the major, phenolic product and the minor, peroxynitrite-specific nitrated product of probe oxidation.
硼酸探针的发展使得对过氧化氢和过氧亚硝酸盐能够进行可靠的检测和定量分析。然而,主要产物是探针的硼酸部分被羟基取代,这两种氧化剂都会产生这种情况。在此,我们描述了线粒体靶向苯基硼酸的邻位异构体如何用于检测和区分过氧亚硝酸盐依赖性和非过氧亚硝酸盐依赖性的探针氧化。该方法突出了探针氧化的主要酚类产物和次要的过氧亚硝酸盐特异性硝化产物的检测和定量。