IDENTIFICATION OF CASPASE-2 SUBSTRATES

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Caspase-2 is the most conserved caspase across different species. It has been reported that Caspase-2 is involved in oocyte death, DNA damage, heat shock-induced death and mitotic catastrophe. However, the exact role of caspase-2 in these biological events is not clear yet due to the lack of knowledge of the substrates of caspase-2. My project is to identify caspase-2 substrates using the established "degradomics" techniques. First, I will treat healthy Jurkat extracts with recombinant active caspase-2. Protein N-terminus in the sample will be labeled, enriched and sequenced by mass spectrometry to identify caspase-2 cleavage. Second, I will use a caspase-2 inducible system to induce endogenous substrate cleavage. Cell extracts will also be labeled, enriched and sequenced by mass spectrometry. Data from both experiments will be compared and integrated to identify caspase-2 substrates.