PRM validation of phosphoproteomic and proteomic analysis of early epileptogenesis in mice at 4 and 24 h post-status epilepticus

The phosphorylation-based signalling and protein changes occurring in the early phases after a pathophysiological insult, like status epilepticus (SE), have not been detailed. In a companion project, the hippocampi of mice treated with pilocarpine and diazepam were examined by tandem mass tag (TMT11plex) mass spectrometry at 4 and 24 h post-status epilepticus (PXD038241). In the accompanying article, the results implicated posttranscriptional regulatory proteins as early targets of increased phosphorylation. Also, the major targets of decreased phosphorylation at 4 h and 24 h were a subset of post synaptic density scaffold proteins, ion channels and neurotransmitter receptors. Here, the earlier work is repeated on protein and phosphorylation site targets representative of the important SE-dependent changes using parallel reaction monitoring (PRM), supporting the main findings.