Global Identification of Caspase Substrates Using PROTOMAP (Protein Topography and Migration Analysis Platform)
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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.
Key wordsSDS-PAGE In-gel digestion Mass spectrometry Spectral counting PROTOMAP Peptograph Substrate identification
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