使用激酶敲除突变体筛选激酶底物。

Screening of kinase substrates using kinase knockout mutants.

作者信息

Umezawa Taishi

机构信息

Faculty of Agriculture, Tokyo University of Agriculture and Technology, 2-24-16 Nakacho, Koganei, Tokyo, 184-8588, Japan,

出版信息

Methods Mol Biol. 2015;1306:59-69. doi: 10.1007/978-1-4939-2648-0_4.

DOI:10.1007/978-1-4939-2648-0_4

PMID:25930693

Abstract

Protein kinases are widely known to be major regulators of various signaling processes, particularly in eukaryotes, including plants. To understand their role in signal transduction pathways, it is necessary to determine which proteins are phosphorylated by these enzymes. Recent studies have applied a comparative phosphoproteomic approach to identify protein kinase substrates in plants. The results demonstrated that kinase knockout mutants are useful for screening protein kinase substrates via such a comparative analysis. Here some technical points are described for the experimental design and comparative analysis using kinase knockout mutants.

摘要

众所周知，蛋白激酶是各种信号传导过程的主要调节因子，尤其是在包括植物在内的真核生物中。为了了解它们在信号转导途径中的作用，有必要确定哪些蛋白质被这些酶磷酸化。最近的研究应用了一种比较磷酸蛋白质组学方法来鉴定植物中的蛋白激酶底物。结果表明，激酶敲除突变体可用于通过这种比较分析筛选蛋白激酶底物。这里描述了一些使用激酶敲除突变体进行实验设计和比较分析的技术要点。