Analysis of protein phosphorylation: methods and strategies for studying kinases and substrates

被引：71

作者：

Peck, SC ^{[1
]}

机构：

[1] Univ Missouri, Columbia, MO 65211 USA

来源：

PLANT JOURNAL | 2006年 / 45卷 / 04期

关键词：

signal transduction; in vitro kinase assay; phosphoproteome; phosphorylation site analysis; phosphatase assay; mitogen-activated protein kinase;

D O I：

10.1111/j.1365-313X.2005.02613.x

中图分类号：

Q94 [植物学];

学科分类号：

071001 ;

摘要：

Protein phosphorylation is a highly conserved mechanism for regulating protein function, being found in all prokaryotes and eukaryotes examined. Phosphorylation can alter protein activity or subcellular localization, target proteins for degradation and effect dynamic changes in protein complexes. In many cases, different kinases may be involved in each of these processes for a single protein, allowing a large degree of combinatorial regulation at the post-translational level. Therefore, knowing which kinases are activated during a response and which proteins are substrates is integral to understanding the mechanistic regulation of a wide range of biological processes. In this paper, I will describe methods for monitoring kinase activity, investigating kinase-substrate specificity, examining phosphorylation in planta and the determination of phosphorylation sites in a protein. In addition, strategic considerations for experimental design and variables will be discussed.

引用

收藏

页码：512 / 522

页数：11

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