Puri Anindita, Tripathi Vasvi, Das Ranabir
National Center for Biological Sciences, Tata Institute of Fundamental Research, Bangalore, India.
Methods Mol Biol. 2025;2957:159-168. doi: 10.1007/978-1-0716-4710-3_11.
Post-translational modifications, such as SUMOylation, play a crucial role in regulating various cellular processes. Traditionally, in vitro SUMOylation assays are carried out using techniques like SDS-PAGE gels and Western blotting, which are often time consuming. These informative methods, which quantitate the increase in substrate's molecular weight, can introduce potential errors and thus a need arises for a more efficient and accurate alternative. We propose a real-time detection method for SUMOylation that utilizes a fluorescence-tagged substrate, which has been recently developed by our group (Tripathi and Das, CP Protein Sci 101:e111, 2020). We believe this method has the potential to significantly enhance the efficiency and accuracy of SUMOylation assessment, thereby advancing our understanding of SUMO signaling and its function.
翻译后修饰，如小泛素样修饰（SUMOylation），在调节各种细胞过程中起着关键作用。传统上，体外SUMOylation分析是使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）凝胶和蛋白质免疫印迹等技术进行的，这些技术通常很耗时。这些通过定量底物分子量增加来提供信息的方法可能会引入潜在误差，因此需要一种更高效、准确的替代方法。我们提出了一种用于SUMOylation的实时检测方法，该方法利用了一种荧光标记的底物，这是我们小组最近开发的（Tripathi和Das，《CP蛋白质科学》101：e111，2020）。我们相信这种方法有可能显著提高SUMOylation评估的效率和准确性，从而增进我们对SUMO信号及其功能的理解。
