Targeted Profiling of Protein Phosphorylation in Plants.

Proteins are crucial for controlling different cellular processes by perceiving and converting external environmental cues into cellular responses. Therefore, regulation of protein activities is pivotal for the development and survival of an organism. This is often mediated by posttranslational modifications, which usually are carried out on specific residues of a target protein by a "writer" protein. The (reversible) modifications of different residues may lead to different signaling outputs. In the case of protein phosphorylation, one of the most common posttranslational modifications, this writer protein is a protein kinase. In this chapter, we report a comprehensive and versatile workflow to identify the phosphorylation profile of a target protein in plants from a putative kinase-target pair by combining an in planta phosphorylation assay and mass spectrometry analysis.