Abstract
Kinase assay linked phosphoproteomics (KALIP) and universal phosphoproteomics (UP) workflows are mass spectrometry (MS)-based phosphoproteomics technologies utilized for studying abiotic stress signaling networks. KALIP consists of executing isotope-labeled phosphorylation reactions in vitro and screening in vivo phosphorylated peptides/pools of substrates. For UP, guanidine hydrochloride (GdnHCl) is used to break down the cell wall before methanol-chloroform precipitation is applied to remove interferences. Both strategies conclude with subsequent analyses of enriched phosphopeptides by complementary MS techniques to determine phosphopeptide abundance changes and protein phosphorylation sites to uncover novel signaling components in the pathway.
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