Department of Biotechnology, University of Verona, Strada le Grazie 15, 37134 Verona, Italy.
Department of Experimental Oncology, European Institute of Oncology (IEO) IRCCS, Via Adamello 16, 20139 Milano, Italy.
J Chromatogr A. 2022 Aug 16;1678:463352. doi: 10.1016/j.chroma.2022.463352. Epub 2022 Jul 19.
Post-translational modifications (PTMs) occur during or after protein biosynthesis and increase the functional diversity of proteome. They comprise phosphorylation, acetylation, methylation, glycosylation, ubiquitination, sumoylation (among many other modifications), and influence all aspects of cell biology. Mass-spectrometry (MS)-based proteomics is the most powerful approach for PTM analysis. Despite this, it is challenging due to low abundance and labile nature of many PTMs. Hence, enrichment of modified peptides is required for MS analysis. This review provides an overview of most common PTMs and a discussion of current enrichment methods for MS-based proteomics analysis. The traditional affinity strategies, including immunoenrichment, chromatography and protein pull-down, are outlined together with their strengths and shortcomings. Moreover, a special attention is paid to chemical enrichment strategies, such as capture by chemoselective probes, metabolic and chemoenzymatic labelling, which are discussed with an emphasis on their recent progress. Finally, the challenges and future trends in the field are discussed.
蛋白质翻译后的修饰（PTMs）发生在蛋白质生物合成过程中或之后，增加了蛋白质组的功能多样性。它们包括磷酸化、乙酰化、甲基化、糖基化、泛素化、SUMO 化（以及许多其他修饰），并影响细胞生物学的各个方面。基于质谱（MS）的蛋白质组学是 PTM 分析最强大的方法。尽管如此，由于许多 PTMs 的丰度低和不稳定，这仍然具有挑战性。因此，需要对修饰肽进行富集，以便进行 MS 分析。本综述概述了最常见的 PTM，并讨论了当前基于 MS 的蛋白质组学分析的富集方法。传统的亲和策略，包括免疫富集、色谱和蛋白质下拉，与它们的优缺点一起概述。此外，特别关注化学富集策略，例如化学选择性探针的捕获、代谢和化学酶标记，讨论重点是其最新进展。最后，讨论了该领域的挑战和未来趋势。
