Bonilla Leo, Means Gary, Lee Kimberly, Patterson Scott
Molecular Sciences, Amgen Inc., Seattle, WA, USA.
Biotechniques. 2008 Apr;44(5):671-9. doi: 10.2144/000112800.
The importance of the analysis of signaling pathways has been proven for many years by the elucidation of key signaling molecules. However, in most cases these pathways tend to represent a rather narrow view of the biological state under investigation. Clearly a more detailed understanding of the complexities of cross-talk between signaling pathways is required to further our knowledge of normal and disease processes. The tools that provide the framework for this increased understanding of biology, those that enable identification, characterization, and quantitation of sites of phosphorylation in proteins, have advanced over the past 25 years. This review will present a brief overview of the history of the tools used in phosphorylation analysis and the latest technologies that are being applied in this field, such as mass spectrometry (for broad-based discovery efforts) and flow cytometry (for translation to clinical applications).
多年来，通过对关键信号分子的阐释，信号通路分析的重要性已得到证实。然而，在大多数情况下，这些通路往往代表了对所研究生物状态的一种相当狭隘的观点。显然，需要更详细地了解信号通路之间复杂的相互作用，以增进我们对正常和疾病过程的认识。在过去25年里，为更深入理解生物学提供框架的工具，即那些能够识别、表征和定量蛋白质磷酸化位点的工具，已经取得了进展。本综述将简要概述磷酸化分析中所用工具的历史以及该领域正在应用的最新技术，如质谱（用于广泛的发现工作）和流式细胞术（用于转化为临床应用）。
