Figure 2.
Caspase substrates identification using proteomics approaches. Two main methods are used to identify caspase substrates: the forward and reverse techniques. The forward method involves treating a cell population with an inducer to trigger a biological process that activates endogenous caspases, such as cell death or cell differentiation. Cells are lysed, and cleaved products either captured or separated for identification using mass spectrometry. In the reverse method, one generates a cell lysate in which endogenous proteases are inhibited and adds a purified caspase of interest. After a suitable time(s), proteolytic fragments are captured, identified, and quantified using mass spectrometry