Activity-based Protein Profiling

Displaying the active proteome

To monitor the manipulation of host enzymes during infection, we exploit activity-based protein profiling (ABPP). ABPP is based on the use of small molecule probes that react with the active sites of enzymes in an activity-dependent manner. Labeling is covalent and irreversible and this facilitates the display of labeled proteins on protein gels and the identification of the labeled proteins by mass spectrometry. There are hundreds of probes reported for different enzyme families. We validate these probes and develop novel probes to extend the range of enzymes we can monitor. Our pioneering work introducing ABPP in plant science has resulted in a large and unique probe library and extensive knowledge on profiling active enzymes. With probes for cysteine, serine and metallo proteases, the proteasome, lipases, esterases, glycosidases, ATP binding proteins, cytochrome P450s and glutathione transferases we have build a powerful platform for international collaborations by hosting visiting scientists. We have extended chemical proteomics in plant science further by establishing reactivity-based protein profiling (RBPP) using probes that label solvent-exposed residues in proteomes.