Martínez-Ruiz Antonio, Lamas Santiago
Centro de Investigaciones Biologicas, Consejo Superior de Investiaciones Cientificas, Rmeira de Maeztu, Madrid.
Methods Enzymol. 2005;396:131-9. doi: 10.1016/S0076-6879(05)96013-8.
Nitric oxide performs some of its direct effects by the induction of various protein posttranslational modifications. Among them, S-nitrosylation of cysteines has gained increasing attention and has been postulated as a signaling mechanism. However, there are still many technical limitations for the detection and identification of this posttranslational modification in cell proteins, and some of them are directly related to the lability of the nitrosothiol bond. We describe protocols for applying a derivatization technique, the biotin switch, which substitutes the S-nitrosylation for a biotinylation in the detection and proteomic identification of S-nitrosylated proteins in intact cells.
一氧化氮通过诱导各种蛋白质翻译后修饰来发挥其一些直接作用。其中，半胱氨酸的S-亚硝基化受到越来越多的关注，并被假定为一种信号传导机制。然而，在细胞蛋白质中检测和鉴定这种翻译后修饰仍存在许多技术限制，其中一些与亚硝基硫醇键的不稳定性直接相关。我们描述了应用一种衍生化技术——生物素开关的方案，该技术在完整细胞中S-亚硝基化蛋白质的检测和蛋白质组学鉴定中，将S-亚硝基化替换为生物素化。
