Rodríguez-Ruiz Marta, Mioto Paulo T, Palma José M, Corpas Francisco J
Department of Biochemistry, Cell and Molecular Biology of Plants, Estación Experimental del Zaidín, Spanish National Research Council (CSIC), C/Profesor Albareda, Granada, Spain.
Bio Protoc. 2017 Sep 20;7(18):e2559. doi: 10.21769/BioProtoc.2559.
In plant cells, the analysis of protein -nitrosothiols (SNOs) under physiological and adverse stress conditions is essential to understand the mechanisms of Nitric oxide (NO)-based signaling. We adapted a previously reported protocol for detecting protein SNOs in animal systems ( King , 2005 ) for plant samples. Briefly, proteins from plant samples are separated via non-reducing SDS-PAGE, then the NO bound by -nitrosylated proteins is released using UV light and, finally, the NO is detected using the fluorescent probe DAF-FM (Rodriguez-Ruiz , 2017). Thus, the approach presented here provides a relatively quick and economical procedure that can be used to compare protein SNOs content in plant samples and provide insight in NO-based signaling in plants.
在植物细胞中，分析生理和逆境胁迫条件下的蛋白质亚硝基硫醇（SNOs）对于理解基于一氧化氮（NO）的信号传导机制至关重要。我们采用了先前报道的用于检测动物系统中蛋白质SNOs的方法（King，2005年）来处理植物样品。简而言之，植物样品中的蛋白质通过非还原SDS-PAGE进行分离，然后使用紫外光释放与亚硝基化蛋白质结合的NO，最后使用荧光探针DAF-FM检测NO（Rodriguez-Ruiz，2017年）。因此，本文介绍的方法提供了一种相对快速且经济的程序，可用于比较植物样品中蛋白质SNOs的含量，并深入了解植物中基于NO的信号传导。
