Chemical Proteomics, Plant Physiology.
Activities of extracellular (apoplastic) proteases and other enzymes during plant-microbe interactions change upon infection with pathogens, partly because the host releases an immune response that includes the oxidative burst and alkalinisation of the apoplast.
To map the effect of these events on the activities of proteases and other secreted proteins, I am applying activity-based protein profiling (ABPP) to detect protein activities that change upon the oxidative burst. In addition, I am establishing hyper-reactive cysteine profiling (HRCP) to identify cysteine residues in proteins that sense the oxidative burst. Both ABPP and HRCP are based on the use of chemical probes that react covalently and irreversibly with proteins, facilitating the detection and identification of the labeled proteins and labeling sites. I further focus on proteins in the apoplast of Nicotiana benthamiana infected with Pseudomonas syringae that both change in activity (by ABPP) and have differential hyper-reactive Cys residues (by HRCP).
Morimoto, K., and Van der Hoorn, R. A. L. (2016) The increasing impact of activity-based protein profiling in plant science. Plant Cell Physiol. 57: 446-461.
Mogami, J., Fujita, Y., Yoshida, T., Tsukiori, Y., Nakagami, H., Nomura, Y., Fujiwara, T., Nishida, S., Yanagisawa, S., Ishida, T., Takahashi, F., Morimoto, K., Kidokoro, S., Mizoi, J., Shinozaki, K., Yamaguchi-Shinozaki, K.(2015) Two Distinct Families of Protein Kinases Are Required for Plant Growth under High External Mg2+ Concentrations in Arabidopsis. Plant Physiol. 167(3): 1039-57.
Sato, H., Mizoi, J., Tanaka, H., Maruyama, K., Qin, F., Osakabe, Y., Morimoto, K., Ohori, T., Kusakabe, K., Nagata, M., Shinozaki, K., Yamaguchi-Shinozaki K. (2014) Arabidopsis Dpb3-1, a DREB2A interactor, specifically enhances heat stress-induced gene expression by forming a heat stress-specific transcriptional complex with NF-Y subunits. Plant Cell. 26(12): 4954-4973.
Morimoto, K., Mizoi, J., Qin, F., Kim, J-S., Sato, H., Osakabe, Y., Shinozaki, K., Yamaguchi-Shinozaki, K. (2013) Stabilisation of Arabidopsis DREB2A is required but not sufficient for induction of its target genes under stress conditions. PloS One. 8(12): e80457.
Qin, F., Kodaira, K., Maruyama, K., Mizoi, J., Tran, L.-S. P, Fujita, Y., Morimoto, K., Shinozaki, K. and Yamaguchi-Shinozaki, K. (2011) SPINDLY, a Negative Regulator of Gibberellic Acid Signaling, Is Involved in the Plant Abiotic Stress Response. Plant Physiol. 157: 1900-1913.