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. Recent advances in the areas of molecular biology and biochemistry are presenting new possibilities for reaching an unprecedented depth and a proteome-wide understanding of phosphorylation processes in plants as well as in other species.
The major goal of Plant Kinases: Methods and Protocols is to provide the experimentalist with a detailed account of the practical steps necessary for successfully carrying out each protocol in his or her own laboratory. Plant protein kinases specifically addressed in this volume are members of the plant MAP kinase cascade, cyclin- and Calcium-dependent protein kinases, and plant sensor and receptor kinases. Written in the highly successful Methods in Molecular Biology series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.
Authoritative and accessible, Plant Kinases: Methods and Protocols will prove a useful laboratory companion to both novice and seasoned researchers by facilitating the practical work that will lead them to new and exciting insights in this dynamic field.
1. Guide to the Book Plant Kinases Nico Dissmeyer and Arp Schnittger
2. The Age of Protein Kinases Nico Dissmeyer and Arp Schnittger
3. Expression and Purification of Active Protein Kinases from Wheat Germ Extracts Boglarka Sonkoly, Viola Bardoczy, and Tamas Meszaros
4. Measurement of Plant Cyclin-Dependent Kinase Activity Using Immunoprecipitation-Coupled and Affinity Purification-Based Kinase Assays and the Baculovirus Expression System Hirofumi Harashima and Masami Sekine
5. Mitogen-Activated Protein Kinase Activity and Reporter Gene Assays in Plants Robert Doczi, Elizabeth Hatzimasoura, and Laszlo Bogre
6. Use of Phospho-site Substitutions to Analyze the Biological Relevance of Phosphorylation Events in Regulatory Networks Nico Dissmeyer and Arp Schnittger
7. Bacterial Assay to Study Plant Sensor Histidine Kinases Lukas Spichal
8. Substrate Analysis of Arabidopsis PP2C-type Protein Phosphatase Julija Umbrasaite, Alois Schweighofer and Irute Meskiene
9. Modulating and Monitoring MAPK Activity During Programmed Cell Death in Pollen Shutian Li and Vernonica E. Franklin-Tong
10. Sensitizing Plant Protein Kinases to Specific Inhibition by ATP-Competitive Molecules Dor Salomon, Chao Zhang, Kevan M. Shokat, and Guido Sessa
11. Fluorescence Fluctuation Analysis of Receptor Kinase Dimerization Mark A. Hink, Sacco C. de Vries, and Antonie J.W.G. Visser
12. Quantifying Degradation Rates of Trans-Membrane Receptor Kinases Niko Geldner
13. Fluorescence Correlation Spectroscopy and Fluorescence Recovery after Photobleaching to Study Receptor Kinase Mobility in planta Mark A.C.J. Kwaaitaal, Marieke Schor, Mark A. Hink, Antonie J.W.G. Visser, and Sacco C. de Vries
14. Bimolecular-Fluorescence Complementation Assay to Monitor Kinase-Substrate Interactions in vivo Stefan Pusch, Nico Dissmeyer, and Arp Schnittger
15. Chemical Genetic Analysis of Protein Kinase Function in Plants Mike Bohmer, Michael Bolker, and Tina Romeis
16. Modified Metal Oxide Affinity Enrichment Combined with 2D-PAGE and Analysis of Phosphoproteomes Thomas Colby, Horst Rohrig, Anne Harzen, and Jurgen Schmidt
17. Phosphoproteomics Using iTRAQ Alexandra M.E. Jones and Thomas S. Nuhse