title: Proteomic discovery of protease substrates.
abstract: Elucidation of in vivo substrate degradomes of a protease is a daunting endeavor because of the large number of proteins in a proteome and often minute and transient amounts of key substrates. Proteomic substrate screens for proteases are currently experiencing impressive progress. Mass spectrometry-based global proteome analysis, interfaced with liquid-chromatography and together with stable isotope labeling strategies, has provided increased coverage and sensitivity for quantitative proteomics. ICAT and iTRAQ labeling have been used to identify a plethora of new matrix metalloproteinase substrates. Emerging techniques focus on the quantitative analysis of proteolytically generated neo amino-termini, which we call terminopes, on a system-wide basis. In vivo SILAC pulse-chase experiments have also enabled the study of individual protein turnover and global proteome dynamics in cells and whole organisms. Together with activity-based probes for the profiling of functional proteases, there is now in place an array of complementary technologies to dissect the 'protease web' and its distortion in pathology.
authors.affiliation: The UBC Centre for Blood Research, Departments of Oral Biological & Medical Sciences, and Biochemistry and Molecular Biology, University of British Columbia, Vancouver, Canada.
authors.forename: Oliver
authors.initials: O
authors.lastname: Schilling
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Overall
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods; D007202:Indicators and Reagents; D007553:Isotope Labeling; D008954:Models, Biological; D010447:Peptide Hydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics / Q000379:methods* / Q000639:trends; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013379:Substrate Specificity
chemical_list: D007202:Indicators and Reagents; D010447:Peptide Hydrolases