Lang Valérie, Aillet Fabienne, Da Silva-Ferrada Elisa, Xolalpa Wendy, Zabaleta Lorea, Rivas Carmen, Rodriguez Manuel S
Inbiomed, Ubiquitylation and Cancer Molecular Biology Laboratory, Cancer Unit Mikeletegi 81, 20009 San Sebastián-Donostia, Spain.
Dpt Biología Molecular y Celular, Centro Nacional de Biotecnología-CSIC, Darwin 3, Madrid 28049, Spain; Centro de Investigación en Medicina Molecular y Enfermedades Crónicas (CIMUS), Universidad de Santiago de Compostela, Instituto de Investigaciones Sanitarias (IDIS), Santiago de Compostela E15706, Spain.
Methods. 2015 May;77-78:112-8. doi: 10.1016/j.ymeth.2014.09.001. Epub 2014 Sep 16.
The function of the tumour suppressor phosphatase and tensin homolog deleted on chromosome 10 (PTEN) is tightly controlled by post-translational modifications (PTMs) including ubiquitin or Small Ubiquitin-related MOdifiers (SUMO). It is known that SUMOylation by SUMO-1, SUMO-2/-3, mono- or polyubiquitylation have a distinct impact on PTEN activity, localisation and/or stability, however the molecular mechanisms governing these processes are still unclear. Studying PTM regulated events has always been a difficult task due to their labile nature. Here, we propose an update on the role of these PTMs on PTEN function, as well as a methodological overview on the use of molecular traps named SUMO Binding Entities (SUBEs) or Tandem Ubiquitin Binding Entities (TUBEs) to capture SUMOylated or Ubiquitylated forms of PTEN respectively. When combined with in vitro SUMOylation or Ubiquitylation assays, the use of molecular traps facilitate the detection of modified forms of PTEN. SUMO and ubiquitin-traps are also suitable to capture endogenously modified forms of PTEN after expression of E3 ligases or treatment with chemical inhibitors. This versatile approach represents an interesting alternative to explore PTEN regulation by SUMO and ubiquitin under physiological or pathological conditions.
10号染色体缺失的磷酸酶和张力蛋白同源物（PTEN）作为一种肿瘤抑制因子，其功能受包括泛素或小泛素相关修饰物（SUMO）在内的翻译后修饰（PTM）严格调控。已知SUMO-1、SUMO-2/-3介导的SUMO化修饰、单泛素化或多泛素化对PTEN的活性、定位和/或稳定性有不同影响，然而，调控这些过程的分子机制仍不清楚。由于PTM的不稳定性，研究PTM调控的事件一直是一项艰巨的任务。在此，我们对这些PTM在PTEN功能中的作用进行了更新，并概述了一种名为SUMO结合实体（SUBE）或串联泛素结合实体（TUBE）的分子陷阱的使用方法，分别用于捕获SUMO化或泛素化形式的PTEN。当与体外SUMO化或泛素化检测相结合时，分子陷阱的使用有助于检测PTM修饰形式。SUMO和泛素陷阱也适用于在E3连接酶表达或化学抑制剂处理后捕获内源性修饰的PTEN形式。这种通用方法是在生理或病理条件下探索SUMO和泛素对PTEN调控的一种有趣替代方法。
