Screening of kinase substrates using kinase knockout mutants.
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.
Faculty of Agriculture, Tokyo University of Agriculture and Technology, 2-24-16 Nakacho, Koganei, Tokyo, 184-8588, Japan, taishi@cc.tuat.ac.jp.
Taishi
T
Umezawa
D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism; D055786:Gene Knockout Techniques; D017343:Genes, Plant; D009154:Mutation; D010766:Phosphorylation; D011494:Protein Kinases / Q000235:genetics* / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D015398:Signal Transduction; D013379:Substrate Specificity
D029681:Arabidopsis Proteins; D011494:Protein Kinases