Author:

Keywords:

Science & Technology, Life Sciences & Biomedicine, Biochemistry & Molecular Biology, Immunology, Microbiology, ACTIVITY-BASED PROBES, AMINO-ACIDS, PRECLINICAL EVALUATION, SUBSTRATE-SPECIFICITY, SELECTIVE INHIBITORS, CYSTEINE PROTEASES, CLEAVABLE LINKER, SERINE PROTEASES, HIGH-THROUGHPUT, ACTIVATION, Animals, Endopeptidases, Enzyme Assays, Humans, Molecular Probe Techniques, Peptide Hydrolases, 0605 Microbiology, 1107 Immunology, 1108 Medical Microbiology, 3107 Microbiology, 3204 Immunology

Abstract:

The activity of proteases is tightly regulated, and dysregulation is linked to a variety of human diseases. For this reason, ABPP is a well-suited method to study protease biology and the design of protease probes has pushed the boundaries of ABPP. The development of highly selective protease probes is still a challenging task. After an introduction, the first section of this chapter discusses several strategies to enable detection of a single active protease species. These range from the usage of non-natural amino acids, combination of probes with antibodies, and engineering of the target proteases. A next section describes the different types of detection tags that facilitate the read-out possibilities including various types of imaging methods and mass spectrometry-based target identification. The power of protease ABPP is illustrated by examples for a selected number of proteases. It is expected that some protease probes that have been evaluated in animal models of human disease will find translation into clinical application in the near future.