title: Activity-based profiling of proteases.
abstract: Proteolytic enzymes are key signaling molecules in both normal physiological processes and various diseases. After synthesis, protease activity is tightly controlled. Consequently, levels of protease messenger RNA and protein often are not good indicators of total protease activity. To more accurately assign function to new proteases, investigators require methods that can be used to detect and quantify proteolysis. In this review, we describe basic principles, recent advances, and applications of biochemical methods to track protease activity, with an emphasis on the use of activity-based probes (ABPs) to detect protease activity. We describe ABP design principles and use case studies to illustrate the application of ABPs to protease enzymology, discovery and development of protease-targeted drugs, and detection and validation of proteases as biomarkers.
authors.affiliation: Departments of 1Chemical and Systems Biology.
authors.forename: Laura E
authors.initials: LE
authors.lastname: Sanman
authors.forename: Matthew
authors.initials: M
authors.lastname: Bogyo
mesh_terms: D000818:Animals; D001671:Biochemistry / Q000379:methods; D015415:Biomarkers / Q000737:chemistry; D020169:Caspases / Q000737:chemistry; D002626:Chemistry, Pharmaceutical / Q000379:methods; D015195:Drug Design; D004926:Escherichia coli / Q000201:enzymology; D006801:Humans; D010447:Peptide Hydrolases / Q000737:chemistry* / Q000502:physiology*; D010455:Peptides / Q000737:chemistry; D040901:Proteomics / Q000379:methods; D013379:Substrate Specificity
chemical_list: D015415:Biomarkers; D010455:Peptides; D010447:Peptide Hydrolases; D020169:Caspases
keywords: activity-based probes; affinity handle; fluorescent imaging; mass spectrometry; proteomics