Abstract
Delineation of the natural substrate scope of proteases is important for understanding the functions of proteolytic pathways in physiology and disease. Herein we describe the protocol for PROTOMAP, a technique that combines SDS-PAGE with tandem mass spectrometry to globally identify shifts in protein migration indicative of proteolytic processing. When applied to cells undergoing apoptosis, this unbiased global method provides a snapshot of the topography and magnitude of proteolytic events associated with programmed cell death.
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References
Dix MM, Simon GM, Cravatt BF (2008) Global mapping of the topography and magnitude of proteolytic events in apoptosis. Cell 134:679–691
Simon GM, Dix MM, Cravatt BF (2009) Comparative assessment of large-scale proteomic studies of apoptotic proteolysis. ACS Chem Biol 4:401–408
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Dix, M.M., Simon, G.M., Cravatt, B.F. (2014). Global Identification of Caspase Substrates Using PROTOMAP (Protein Topography and Migration Analysis Platform). In: V. Bozhkov, P., Salvesen, G. (eds) Caspases,Paracaspases, and Metacaspases. Methods in Molecular Biology, vol 1133. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0357-3_3
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DOI: https://doi.org/10.1007/978-1-4939-0357-3_3
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-0356-6
Online ISBN: 978-1-4939-0357-3
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