Xu Guoqiang, Deglincerti Alessia, Paige Jeremy S, Jaffrey Samie R
Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and College of Pharmaceutical Sciences, Soochow University, Suzhou, Jiangsu, China.
Methods Mol Biol. 2014;1174:57-71. doi: 10.1007/978-1-4939-0944-5_4.
Protein ubiquitination plays critical roles in many biological processes. However, functional studies of protein ubiquitination in eukaryotic cells are limited by the ability to identify protein ubiquitination sites. Unbiased high-throughput screening methods are necessary to discover novel ubiquitination sites that play important roles in cellular regulation. Here, we describe an immunopurification approach that enriches ubiquitin remnant-containing peptides to facilitate downstream mass spectrometry (MS) identification of lysine ubiquitination sites. This approach can be utilized to identify ubiquitination sites from proteins in a complex mixture.
蛋白质泛素化在许多生物学过程中发挥着关键作用。然而，真核细胞中蛋白质泛素化的功能研究受到识别蛋白质泛素化位点能力的限制。为了发现细胞调节中发挥重要作用的新型泛素化位点，无偏倚的高通量筛选方法是必要的。在此，我们描述了一种免疫纯化方法，该方法富集含泛素残基的肽段，以促进赖氨酸泛素化位点的下游质谱（MS）鉴定。这种方法可用于从复杂混合物中的蛋白质识别泛素化位点。
