Abstract
Activity-based protein profiling (ABPP) is a powerful analytical method to detect and compare the activity of proteins in proteomes. This is achieved using specific activity-based probes that are often derived from inhibitors and are linked to reporter groups like rhodamine or biotin for fluorescence detection and/or affinity purification, respectively. The probes react with the active site residue of proteins and become covalently and irreversibly attached, facilitating the separation, detection and identification of the labelled proteins. In this protocol we describe all the steps required for labelling, purification and identification of labelled proteins from gels and show how activities in two proteomes can be compared. The identification of serine hydrolases from Arabidopsis plants infected with Botrytis cinerea using the trifunctional probe TriFP is used as an example.
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Acknowledgements
This work was supported by the Max Planck Society and the Deutsche Forschungsgemeinschaft projects HO3983/3-1,2 and HO3983/4-1
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Kaschani, F., Gu, C., van der Hoorn, R.A.L. (2012). Activity-Based Protein Profiling of Infected Plants. In: Bolton, M., Thomma, B. (eds) Plant Fungal Pathogens. Methods in Molecular Biology, vol 835. Humana Press. https://doi.org/10.1007/978-1-61779-501-5_3
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DOI: https://doi.org/10.1007/978-1-61779-501-5_3
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