Activity based proteomics, or activity based protein profiling (ABPP) is a functional proteomic technology that uses specially designed chemical probes that react with mechanistically-related classes of enzymes. The basic unit of ABPP is the probe which typically consists of two elements: a reactive group (RG) and a tag. Additionally, some probes may contain a binding group which enhances selectivity. The reactive group usually contains an electrophile that gets covalently-linked to a nucleophilic residue in the active site of an active enzyme. An enzyme that is inhibited by enzyme inhibitors or post-translational modifications will not react with an activity-based probe. The tag may be either a reporter such as a fluorophore or an affinity label such as biotin or an alkyne or azide for use with the Huisgen 1,3-dipolar cycloaddition (also known as click chemistry).
A major advantage of ABPP is the ability to monitor the availability of the enzyme active site directly, rather than being limited to protein or mRNA abundance. With classes of enzymes such as the serine proteases and metalloproteases that often interact with endogenous inhibitors or that exist as inactive zymogens, this technique offers a valuable advantage over traditional techniques that rely on abundance rather than activity.
Finally, in recent years ABPP has been combined with tandem mass spectrometry enabling the identification of hundreds of active enzymes from a single sample. This technique, known as ABPP-MudPIT (multidimensional protein identification technology) is especially useful for profiling inhibitor selectivity as the potency of an inhibitor can be tested against hundreds of targets simultaneously.
ABPP was originally invented by James C. Powers (Georgia Tech, Atlanta) in the early 1990s and subsequently adopted and developed by the other labs, for example Benjamin Cravatt III at The Scripps Research Institute, Matthew Bogyo at Stanford, and others.