Methods for the proteomic identification of protease substrates.
Proteolysis is a key regulatory post-translational modification in diverse cellular processes including programed cell death, immune function, and development. Tracking proteolytic events has become a focus of researchers assessing the downstream consequences of protease activation. In this review we summarize unbiased methods for identifying protease substrates and tracking the extent of cleavage, a field termed 'degradomics'. These include one-dimensional and two-dimensional gel-based methods for identifying protease substrates, N-terminal peptide identification methods for simultaneously identifying substrates and cleavage sites, and approaches for the quantitation of cleavage events during endogenous proteolysis. Individual methods have identified more than 300 caspase-cleaved targets during apoptosis suggesting broad future applications for these technologies.
Agard
Nicholas J
NJ
Department of Pharmaceutical Chemistry, University of California, San Francisco, UCSF MC 2552, 1700 4th St, San Francisco, CA 94158, United States of America.
Wells
James A
JA
D000818:Animals; D004586:Electrophoresis; D006801:Humans; D010447:Peptide Hydrolases / Q000737:chemistry / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D013379:Substrate Specificity
D010447:Peptide Hydrolases