title: Detection of protease activity in cells and animals.
abstract: Proteases are involved in a wide variety of biologically and medically important events. They are entangled in a complex network of processes that regulate their activity, which makes their study intriguing, but challenging. For comprehensive understanding of protease biology and effective drug discovery, it is therefore essential to study proteases in models that are close to their complex native environments such as live cells or whole organisms. Protease activity can be detected by reporter substrates and activity-based probes, but not all of these reagents are suitable for intracellular or in vivo use. This review focuses on the detection of proteases in cells and in vivo. We summarize the use of probes and substrates as molecular tools, discuss strategies to deliver these tools inside cells, and describe sophisticated read-out techniques such as mass spectrometry and various imaging applications. This article is part of a Special Issue entitled: Physiological Enzymology and Protein Functions.
authors.affiliation: Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, Geert Grooteplein 26-28, 6525 GA Nijmegen, The Netherlands.
authors.forename: Martijn
authors.initials: M
authors.lastname: Verdoes
authors.affiliation: Leibniz Institute for Analytical Sciences, ISAS, e.V., Otto-Hahn-Str. 6b, 44227 Dortmund, Germany; Laboratory of Chemical Biology, Department of Cellular and Molecular Medicine, University of Leuven, Herestraat 49, Box 802, B-3000, Belgium. Electronic address: steven.verhelst@isas.de.
authors.forename: Steven H L
authors.initials: SH
authors.lastname: Verhelst
mesh_terms: D000818:Animals; D055162:Biocatalysis; D020134:Catalytic Domain; D057075:Enzyme Assays / Q000379:methods; D042541:Intracellular Space / Q000201:enzymology*; D008954:Models, Biological; D008958:Models, Molecular; D015335:Molecular Probes / Q000737:chemistry; D010447:Peptide Hydrolases / Q000737:chemistry* / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D015335:Molecular Probes; D010447:Peptide Hydrolases
keywords: Activity-based probes; Degradomics; Imaging; Proteases; Substrate reporters