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Full Description
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.
Contents
Guide to the Book Plant Kinases.- The Age of Protein Kinases.- Expression and Purification of Active Protein Kinases from Wheat Germ Extracts.- Measurement of Plant Cyclin-Dependent Kinase Activity Using Immunoprecipitation-Coupled and Affinity Purification-Based Kinase Assays and the Baculovirus Expression System.- Mitogen-Activated Protein Kinase Activity and Reporter Gene Assays in Plants.- Use of Phospho-site Substitutions to Analyze the Biological Relevance of Phosphorylation Events in Regulatory Networks.- Bacterial Assay to Study Plant Sensor Histidine Kinases.- Substrate Analysis of Arabidopsis PP2C-type Protein Phosphatase.- Modulating and Monitoring MAPK Activity During Programmed Cell Death in Pollen.- Sensitizing Plant Protein Kinases to Specific Inhibition by ATP-Competitive Molecules.- Fluorescence Fluctuation Analysis of Receptor Kinase Dimerization.- Quantifying Degradation Rates of Trans-Membrane Receptor Kinases.- Fluorescence Correlation Spectroscopy and Fluorescence Recovery after Photobleaching to Study Receptor Kinase Mobility in planta.- Bimolecular-Fluorescence Complementation Assay to Monitor Kinase-Substrate Interactions in vivo.- Chemical Genetic Analysis of Protein Kinase Function in Plants.- Modified Metal Oxide Affinity Enrichment Combined with 2D-PAGE and Analysis of Phosphoproteomes.- Phosphoproteomics Using iTRAQ
