Current Developments in Activity-Based Protein Profiling

Abstract

Activity-based protein profiling (ABPP) has emerged as a powerful strategy to study the activity of enzymes in complex proteomes. The aim of ABPP is to selectively visualize only the active forms of particular enzymes using chemical probes termed activity-based probes (ABPs). These probes are directed to the active site of a particular target protein (or protein family) where they react in a mechanism-based manner with an active site residue. This results in the selective labeling of only the catalytically active form of the enzyme, usually in a covalent manner. Besides the monitoring of a specific enzymatic activity, ABPP strategies have also been used to identify and characterize (unknown) protein functions, to study up- and down-regulation of enzymatic activity in various disease states, to discover and evaluate putative new enzyme inhibitors, and to identify the protein targets of covalently binding natural products. In this Topical Review we will provide a brief overview of some of the recent developments in the field of ABPP.

Subfamily-Specific Fluorescent Probes for Cysteine Proteases Display Dynamic Protease Activities during Seed Germination

“…Protease activity profiling (also called activity-based protein profiling of proteases) is an easy and powerful method to monitor the active state of proteases in crude extracts or living organisms (Heal et al, 2011;Serim et al, 2012;Haedke et al, 2013;Willems et al, 2014). Protease activity profiling is based on the use of chemical probes that react covalently with the active site of proteases in an activitydependent manner.…”

Phosphono Bisbenzguanidines as Irreversible Dipeptidomimetic Inhibitors and Activity‐Based Probes of Matriptase‐2

“…[20,21,[33][34][35] The visualization of the target is restricted to the activeform of the enzyme, which is of particular importance,b ecause enzyme expression levels do not alwaysc orrelate with the activity or the activity might be regulated post-translationally. [34,36] These probes consist of three essentialc omponents: ar eactive group, ar ecognition element, and ar eporter tag. The reactive group, as o-called warhead, leads to ac ovalent attachment to an active-site nucleophile.…”

The potential of bioorthogonal chemistry for correlative light and electron microscopy: a call to arms

“…Although genetic modification is needed for this approachwith the same downsides as other genetic techniques-it is a great addition to the bioorthogonal toolkit. Attachment of abiotic tags to covalent enzyme inhibitors allows even to selective visualize active populations of enzymes in a complex mixture [17][18][19]. Moreover, it can be applied to the tagging of biomolecules in living multicellular organisms, such as Caenorhabditis elegans [20], zebrafish [21,22] and mice [23].…”