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Preparation and In Vivo Use of an Activity-based Probe for N-acylethanolamine Acid Amidase
Published on: November 23, 2016
Synthesis and Application of Activity-Based Probes for Serine Proteases
Alba Ramos-Llorca1, Valeria Parravicini1, Koen Augustyns1
1Laboratory of Medicinal Chemistry, Department of Pharmaceutical Sciences, Faculty of Pharmaceutical, Biomedical and Veterinary Sciences, Campus Drie Eiken, University of Antwerp, Antwerp, Belgium.
None:
Activity-based probes (ABPs) are chemical molecules that covalently react with the active form of the targeted enzyme. These probes are useful for target and biomarker identification, as well as drug discovery. This chapter describes the creation of an ABP library tailored to serine proteases, featuring a bioorthogonal tag and a range of diphenyl phosphonate warheads to improve selectivity and reactivity. We detail a protocol for synthesizing substituted warheads, enhancing probe affinity. The effectiveness of these ABPs is validated biochemically through analyses of their inhibitory potencies and kinetic behaviors. Furthermore, we illustrate their application in labeling serine proteases, using fluorescence scanning for detection.
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