UCD School of Medicine, Conway Institute, University College Dublin, Belfield, Dublin 4, Ireland.
J Proteomics. 2022 May 15;259:104558. doi: 10.1016/j.jprot.2022.104558. Epub 2022 Mar 10.
Phosphorylation is a major regulatory mechanism controlling protein and cell function. Phosphoproteomics is continuing to reveal the extent and complexity of protein phosphorylation. In particular, most proteins are emerging to contain multiple phosphorylation sites. However, phosphoproteomics has outpaced current understanding of the functional roles of individual phospho-sites. In this paper the Phos-tag gel method is presented and discussed in the context of other available tools for phosphorylation research. Strengths and weaknesses of Phos-tag gels are outlined and an integrated approach to phosphorylation research is proposed. SIGNIFICANCE: The Phos-tag gel method has unique strengths especially regarding the analysis of multi-site phosphorylation. A combined approach including Phos-tag gels together with other methods like isotope labelling, phospho-specific antibodies, and mass spectrometry is required to advance current understanding of protein phosphorylation.
磷酸化是控制蛋白质和细胞功能的主要调节机制。磷酸蛋白质组学不断揭示蛋白质磷酸化的程度和复杂性。特别是，越来越多的蛋白质被发现含有多个磷酸化位点。然而，磷酸蛋白质组学已经超过了目前对单个磷酸化位点功能作用的理解。本文介绍了 Phos-tag 凝胶法，并结合其他可用的磷酸化研究工具对其进行了讨论。Phos-tag 凝胶的优缺点进行了概述，并提出了一种综合的磷酸化研究方法。意义：Phos-tag 凝胶法具有独特的优势，特别是在分析多磷酸化方面。为了提高对蛋白质磷酸化的理解，需要结合 Phos-tag 凝胶法和其他方法，如同位素标记、磷酸特异性抗体和质谱法。
