Yao Qiuming, Schulze Waltraud X, Xu Dong
Department of Computer Science and Bond Life Sciences Center, University of Missouri, Columbia, MO, 65211, USA.
Methods Mol Biol. 2015;1306:217-28. doi: 10.1007/978-1-4939-2648-0_17.
Protein phosphorylation events on serine, threonine, and tyrosine residues are the most pervasive protein covalent bond modifications in plant signaling. Both low and high throughput studies reveal the importance of phosphorylation in plant molecular biology. Although becoming more and more common, the proteome-wide screening on phosphorylation by experiments remains time consuming and costly. Therefore, in silico prediction methods are proposed as a complementary analysis tool to enhance the phosphorylation site identification, develop biological hypothesis, or help experimental design. These methods build statistical models based on the experimental data, and they do not have some of the technical-specific bias, which may have advantage in proteome-wide analysis. More importantly computational methods are very fast and cheap to run, which makes large-scale phosphorylation identifications very practical for any types of biological study. Thus, the phosphorylation prediction tools become more and more popular. In this chapter, we will focus on plant specific phosphorylation site prediction tools, with essential illustration of technical details and application guidelines. We will use Musite, PhosPhAt and PlantPhos as the representative tools. We will present the results on the prediction of the Arabidopsis protein phosphorylation events to give users a general idea of the performance range of the three tools, together with their strengths and limitations. We believe these prediction tools will contribute more and more to the plant phosphorylation research community.
丝氨酸、苏氨酸和酪氨酸残基上的蛋白质磷酸化事件是植物信号传导中最普遍的蛋白质共价键修饰。低通量和高通量研究均揭示了磷酸化在植物分子生物学中的重要性。尽管实验性的全蛋白质组磷酸化筛选越来越普遍,但仍然耗时且成本高昂。因此,提出了基于计算机的预测方法作为一种补充分析工具,以加强磷酸化位点的识别、提出生物学假设或辅助实验设计。这些方法基于实验数据构建统计模型,不存在某些技术特异性偏差,这在全蛋白质组分析中可能具有优势。更重要的是,计算方法运行速度非常快且成本低廉,这使得大规模磷酸化鉴定对于任何类型的生物学研究都非常实用。因此,磷酸化预测工具越来越受欢迎。在本章中,我们将重点介绍植物特异性磷酸化位点预测工具,并对技术细节和应用指南进行必要说明。我们将使用Musite、PhosPhAt和PlantPhos作为代表性工具。我们将展示对拟南芥蛋白质磷酸化事件的预测结果,让用户对这三种工具的性能范围、优势和局限性有一个大致了解。我们相信这些预测工具将为植物磷酸化研究领域做出越来越大的贡献。