Plant Phosphoproteomics, 2015
Methods and Protocols
Methods in Molecular Biology Series, Vol. 1306

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.

1. The Plant Kinome

            Monika Zulawski and Waltraud X. Schulze

2. Phosphatases in Plants

            Alois Schweighofer and Irute Meskiene

3. Phosphoproteomics in Cereals

            Pingfang Yang

4. Screening of Kinase Substrates Using Kinase-Knockout Mutants

            Taishi Umezawa

5. Phosphopeptide Profiling of Receptor Kinase Mutants

            Xu Na Wu and Waltraud X. Schulze

6. Combining Metabolic ¹⁵N Labelling with Improved TandemMOAC for Enhanced Probing of the Phosphoproteome

            Martin Thomas, Wolfgang Hoehenwarter, Uwe Conrath, and Gerold J.M. Beckers

7. Kinase Activity and Specificity Assay Using Synthetic Peptides

            Xu Na Wu and Waltraud X. Schulze

8. Absolute Quantitation of Protein Post-Translational Modification Isoform

            Zhu Yang and Ning Li

9. Phosphorylation Stoichiometry Determination in Plant Photosynthetic Membranes

            Björn Ingelsson, Rikard Fristedt, and Maria V. Turkina

10. Phosphopeptide Immuno-Affinity Enrichment to Enhance Detection of Tyrosine Phosphorylation in Plants

            Sharon C. Mithoe and Frank L.H. Menke

11. The Peptide Microarray ChloroPhos1.0: A Screening Tool for the Identification of Arabidopsis thaliana Chloroplast Protein Kinase Substrates

            Anna Schönberg and Sacha Baginsky

12. Plant Protein Kinase Substrates Identification Using Protein Microarrays

            Shisong Ma and Savithramma P. Dinesh-Kumar

13. Targeted Analysis of Protein Phosphorylation by 2D Electrophoresis

            Kristin Mayer, Sally Albrecht, and Andreas Schaller

14. Computational Phosphorylation Network Reconstruction: Methods and Resources

            Guangyou Duan and Dirk Walther

15. Computational Identification of Protein Kinases and Kinase-Specific Substrates in Plants

            Han Cheng, Yongbo Wang, Zexian Liu, and Yu Xue

16. Databases for Plant Phosphoproteomics

            Waltraud X. Schulze, Qiuming Yao, and Dong Xu

17. Phosphorylation Site Prediction in Plants

            Qiuming Yao, Waltraud X. Schulze, and Dong Xu

Includes cutting-edge methods and protocols in plant phosphoproteomics

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts