Figure 2. Proposed kinome profiling applications to major depressive disorders and antidepressant treatment.

As described in Duric et al., 2010, cellular extracts can be obtained from post-mortem brain tissue with MDD pathology or long-term antidepressant treatment. Protein fractions are added to a column of sepharose beads covalently linked to a number of different type I kinase inhibitors (adapted from Graves et al., 2013). The active kinases preferentially bind to the inhibitors and remain on the column. The kinases are then eluted from the columns and digested with trypsin. The tryptic peptides are labeled with isobaric tags for relative and absolute quantitation (iTRAQ) and identified and quantitated with mass spectrometry. If a kinase is activated by a specific treatment or pathology, more of that kinase will be bound to the column. Quantity of kinase bound from a treatment over control yields kinome activity profile.