Analytical Signalling Group, Centre for Cell Signalling, Barts Cancer Institute-CR-UK Centre of Excellence, Queen Mary University of London, John Vane Science Centre, Charterhouse Square, London, UK.
Amino Acids. 2012 Sep;43(3):1009-24. doi: 10.1007/s00726-012-1288-9. Epub 2012 Jul 22.
Phosphoproteomics is increasingly used to address a wide range of biological questions. However, despite some success, techniques for phosphoproteomics are not without challenges. Phosphoproteins are present in cells in low abundance relative to their unphosphorylated counterparts; therefore phosphorylated proteins (or phosphopeptides after protein digestion) are rarely detected in standard shotgun proteomics experiments. Thus, extraction of phosphorylated polypeptides from complex mixtures is a critical step in the success of phosphoproteomics experiments. Intense research over the last decade has resulted in the development of powerful techniques for phosphopeptide enrichment prior to analysis by mass spectrometry. Here, we review how the development of IMAC, MOAC, chemical derivatization and antibody affinity purification and chromatography is contributing to the evolution of phosphoproteomics techniques. Although further developments are needed for the technology to reach maturity, current state-of-the-art techniques can already be used as powerful tools for biological research.
磷酸化蛋白质组学越来越多地被用于解决各种生物学问题。然而，尽管取得了一些成功，但磷酸化蛋白质组学技术并非没有挑战。相对于其未磷酸化的对应物，磷酸化蛋白质在细胞中含量较低；因此，在标准的鸟枪法蛋白质组学实验中很少能检测到磷酸化蛋白质（或蛋白质消化后的磷酸肽）。因此，从复杂混合物中提取磷酸化多肽是磷酸化蛋白质组学实验成功的关键步骤。在过去的十年中，人们进行了大量的研究，开发了用于在通过质谱分析之前富集磷酸肽的强大技术。在这里，我们回顾了 IMAC、MOAC、化学衍生化和抗体亲和纯化和色谱法的发展如何促进磷酸化蛋白质组学技术的发展。尽管该技术还需要进一步发展才能达到成熟，但目前最先进的技术已经可以作为生物学研究的有力工具。
