Xu Ping, Peng Junmin
Department of Human Genetics, Center for Neurodegenerative Disease, Emory University, Atlanta, GA 30322, USA.
Biochim Biophys Acta. 2006 Dec;1764(12):1940-7. doi: 10.1016/j.bbapap.2006.09.004. Epub 2006 Sep 14.
Protein modification by ubiquitin is a central regulatory mechanism in eukaryotic cells. Recent proteomics developments in mass spectrometry enable systematic analysis of cellular components in the ubiquitin pathway. Here, we review the advances in analyzing ubiquitinated substrates, determining modified lysine residues, quantifying polyubiquitin chain topologies, as well as profiling deubiquitinating enzymes based on the activity. Moreover, proteomic approaches have been developed for probing the interactome of proteasome and for identifying proteins with ubiquitin-binding domains. Similar strategies have been applied on the studies of the modification by ubiquitin-like proteins as well. These strategies are discussed with respect to their advantages, limitations and potential improvements. While the utilization of current methodologies has rapidly expanded the scope of protein modification by the ubiquitin family, a more active role is anticipated in the functional studies with the emergence of quantitative mass spectrometry.
泛素介导的蛋白质修饰是真核细胞中的一种核心调控机制。近年来，质谱蛋白质组学的发展使得对泛素途径中的细胞成分进行系统分析成为可能。在此，我们综述了在分析泛素化底物、确定修饰的赖氨酸残基、量化多聚泛素链拓扑结构以及基于活性对去泛素化酶进行分析等方面所取得的进展。此外，还开发了蛋白质组学方法来探测蛋白酶体的相互作用组以及鉴定具有泛素结合结构域的蛋白质。类似的策略也已应用于类泛素蛋白修饰的研究。本文讨论了这些策略的优点、局限性及潜在的改进方向。虽然目前方法的应用迅速扩展了泛素家族介导的蛋白质修饰研究范围，但随着定量质谱技术的出现，预计其在功能研究中将发挥更积极的作用。
