Dissmeyer Nico, Schnittger Arp
Leibniz Institute of Plant Biochemistry (IPB), Independent Junior Research Group on Protein Recognition and Degradation, Halle (Saale), Germany.
Methods Mol Biol. 2011;779:7-52. doi: 10.1007/978-1-61779-264-9_2.
Major progress has been made in unravelling of regulatory mechanisms in eukaryotic cells. Modification of target protein properties by reversible phosphorylation events has been found to be one of the most prominent cellular control processes in all organisms. The phospho-status of a protein is dynamically controlled by protein kinases and counteracting phosphatases. Therefore, monitoring of kinase and phosphatase activities, identification of specific phosphorylation sites, and assessment of their functional significance are of crucial importance to understand development and homeostasis. Recent advances in the area of molecular biology and biochemistry, for instance, mass spectrometry-based phosphoproteomics or fluorescence spectroscopical methods, open new possibilities to reach an unprecidented depth and a proteome-wide understanding of phosphorylation processes in plants and other species. In addition, the growing number of model species allows now deepening evolutionary insights into signal transduction cascades and the use of kinase/phosphatase systems. Thus, this is the age where we move from an understanding of the structure and function of individual protein modules to insights how these proteins are organized into pathways and networks. In this introductory chapter, we briefly review general definitions, methodology, and current concepts of the molecular mechanisms of protein kinase function as a foundation for this methods book. We briefly review biochemistry and structural biology of kinases and provide selected examples for the role of kinases in biological systems.
在揭示真核细胞调控机制方面已取得重大进展。通过可逆磷酸化事件改变靶蛋白特性，已被发现是所有生物体中最突出的细胞控制过程之一。蛋白质的磷酸化状态由蛋白激酶和起抵消作用的磷酸酶动态控制。因此，监测激酶和磷酸酶活性、鉴定特定磷酸化位点以及评估其功能意义对于理解发育和体内平衡至关重要。分子生物学和生物化学领域的最新进展，例如基于质谱的磷酸化蛋白质组学或荧光光谱方法，为以前所未有的深度和全蛋白质组范围理解植物及其他物种中的磷酸化过程开辟了新的可能性。此外，越来越多的模式物种使得现在能够更深入地了解信号转导级联反应的进化，并利用激酶/磷酸酶系统。因此，这是一个我们从了解单个蛋白质模块的结构和功能，转向深入了解这些蛋白质如何组织成途径和网络的时代。在这一介绍性章节中，我们简要回顾蛋白质激酶功能分子机制的一般定义、方法和当前概念，作为这本方法书籍的基础。我们简要回顾激酶的生物化学和结构生物学，并提供激酶在生物系统中作用的选定实例。
