Bermúdez-López Marcelino, Aragón Luis
Cell Cycle Group, Medical Research Council (MRC) Clinical Sciences Centre, Imperial College London, Hammersmith Hospital, Du Cane Road, London, W12 0NN, UK.
Methods Mol Biol. 2017;1515:55-64. doi: 10.1007/978-1-4939-6545-8_4.
Cohesin is a protein complex with key roles in chromosome biology, from chromatid segregation to DNA repair. Cohesin function is regulated by several posttranslational modifications, including phosphorylation, acetylation, ubiquitylation, and SUMOylation. Recent studies have shown that cohesin SUMOylation is essential for sister chromatid cohesion during normal cell cycle and in response to DNA damage. Posttranslational modification by the small ubiquitin-like modifier (SUMO) is a field in expansion, however, detecting SUMOylation can be challenging because the amount of modified substrates are usually low and de-conjugation during sample preparation often occurs. In this chapter we describe a method that can be adapted to different model organisms, and substrates to detect SUMOylation. We focus on cohesin and show that SUMOylation indeed occurs in most of the subunits of budding yeast cohesin.
黏连蛋白是一种蛋白质复合体，在染色体生物学中发挥关键作用，从染色单体分离到DNA修复。黏连蛋白的功能受多种翻译后修饰调控，包括磷酸化、乙酰化、泛素化和小泛素样修饰蛋白（SUMO）化。最近的研究表明，黏连蛋白SUMO化对于正常细胞周期以及对DNA损伤作出反应时的姐妹染色单体黏连至关重要。小泛素样修饰蛋白（SUMO）介导的翻译后修饰是一个不断扩展的领域，然而，检测SUMO化可能具有挑战性，因为修饰底物的量通常很低，并且在样品制备过程中经常发生去共轭作用。在本章中，我们描述了一种可适用于不同模式生物和底物以检测SUMO化的方法。我们聚焦于黏连蛋白，并表明SUMO化确实发生在芽殖酵母黏连蛋白的大多数亚基中。
