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This detailed volume addresses recent developments in phosphoproteomic techniques with a particular focus on the plant system. Over the recent decades, proteomic methods were refined to study the significance and dynamics of protein phosphorylation in various biological contexts. However, working with plant tissue imposes particular challenges to the biologist which are attributed to the rigid cell wall making protein extraction more difficult, the skewed protein abundance with Rubisco as a highly abundant protein and a large central vacuole leading to low protein yield and increased degradative enzyme activity. The methodologies in this book seek to move beyond these issues. Written for the Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols and tips on troubleshooting and avoiding known pitfalls.
Practical and authoritative, Plant Phosphoproteomics: Methods and Protocols serves as an ideal reference for researchers investigating this vital area of plant science.
List of contents
The Plant Kinome.- Phosphatases in Plants.- Phosphoproteomics in Cereals.- Screening of Kinase Substrates Using Kinase-Knockout Mutants.- Phosphopeptide Profiling of Receptor Kinase Mutants.- Combining Metabolic 15N Labelling with Improved TandemMOAC for Enhanced Probing of the Phosphoproteome.- Kinase Activity and Specificity Assay Using Synthetic Peptides.- Absolute Quantitation of Protein Post-Translational Modification Isoform.- Phosphorylation Stoichiometry Determination in Plant Photosynthetic Membranes.- Phosphopeptide Immuno-Affinity Enrichment to Enhance Detection of Tyrosine Phosphorylation in Plants.- The Peptide Microarray ChloroPhos1.0: A Screening Tool for the Identification of Arabidopsis thaliana Chloroplast Protein Kinase Substrates.- Plant Protein Kinase Substrates Identification Using Protein Microarrays.- Targeted Analysis of Protein Phosphorylation by 2D Electrophoresis.- Computational Phosphorylation Network Reconstruction: Methods and Resources.- Computational Identification of Protein Kinases and Kinase-Specific Substrates in Plants.- Databases for Plant Phosphoproteomics.- Phosphorylation Site Prediction in Plants.
Summary
This detailed volume addresses recent developments in phosphoproteomic techniques with a particular focus on the plant system. Over the recent decades, proteomic methods were refined to study the significance and dynamics of protein phosphorylation in various biological contexts. However, working with plant tissue imposes particular challenges to the biologist which are attributed to the rigid cell wall making protein extraction more difficult, the skewed protein abundance with Rubisco as a highly abundant protein and a large central vacuole leading to low protein yield and increased degradative enzyme activity. The methodologies in this book seek to move beyond these issues. Written for the Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols and tips on troubleshooting and avoiding known pitfalls.
Practical and authoritative, Plant Phosphoproteomics: Methods and Protocols serves as an ideal reference for researchers investigating this vital area of plant science.
