Identification of Cold-induced MAP Kinase Substrates in Arabidopsis Thaliana through Protein Kinase Assay Linked-Phosphoproteomics

摘要

Mitogen-activated protein kinase (MAPK) cascades are important for amplifying intracellular signal transduction as plant suffering from extracellular stimuli such as hormone, pathogen and cold treatment(1). Despite it has been known that both abiotic and biotic stresses triggers the activation of MAPK3/6 kinase activities, the downstream MPK3/6 substrates regulating individual stress response are still not completely understood. Among these environmental stresses, low temperature is one of the main stresses that plants need to deal with during their life cycle(2), and cold stress is very critical in affecting agricultural product especially in early spring and late autumn. Here we have applied the proteomic strategy, termed stable isotope-labeld protein kinase assay-linked phosphoproteomics (siKALIP)(3), by coupling in vitro kinase assay-based substrates screening with in vivo kinase-dependent quantitative phosphoproteomics. In this study, we aim to identify novel cold-induced MPK6 substrates by proKALIP approach in Arabidopsis Thaliana.