Abbas Kahina, Riquier Sylvie, Drapier Jean-Claude
Institut de Chimie des Substances Naturelles, Centre National de la Recherche Scientifique, Gif-sur-Yvette, France.
Methods Enzymol. 2013;527:113-28. doi: 10.1016/B978-0-12-405882-8.00006-4.
Peroxiredoxins (Prxs) are a family of peroxidases that maintain thiol homeostasis by catalyzing the reduction of organic hydroperoxides, H2O2, and peroxynitrite. Eukaryotic 2-Cys-Prxs, also referred to as typical Prxs, can be inactivated by oxidation of the catalytic cysteine to sulfinic acid, which may regulate the intracellular messenger function of H2O2. A small redox protein, sulfiredoxin (Srx), has been shown to reduce sulfinylated 2-Cys-Prxs and thus to regenerate active 2-Cys-Prxs. We previously reported that cytokine-induced nitric oxide (NO) intervenes in this pathway by decreasing the level of 2-Cys overoxidation and by upregulating Srx through the activation of the transcription factor nuclear factor erythroid 2-related factor (Nrf2). Here, we describe the methods used to monitor the interplay between NO and H2O2 in the regulation of the Prx/Srx system in immunostimulated macrophages, which produce both reactive oxygen species and NO.
过氧化物酶体增殖物激活受体（Prxs）是一类过氧化物酶，通过催化有机氢过氧化物、过氧化氢和过氧亚硝酸盐的还原作用来维持硫醇稳态。真核生物的2-半胱氨酸过氧化物酶（2-Cys-Prxs），也被称为典型的Prxs，可因催化性半胱氨酸氧化为亚磺酸而失活，这可能会调节过氧化氢的细胞内信使功能。一种小的氧化还原蛋白，硫氧还蛋白（Srx），已被证明可还原亚磺酰化的2-Cys-Prxs，从而使活性2-Cys-Prxs再生。我们之前报道，细胞因子诱导的一氧化氮（NO）通过降低2-Cys过度氧化水平并通过激活转录因子核因子红细胞2相关因子（Nrf2）上调Srx，来干预这一途径。在此，我们描述了用于监测免疫刺激巨噬细胞中Prx/Srx系统调控过程中NO与过氧化氢之间相互作用的方法，这些巨噬细胞会产生活性氧和NO。
