Activity-Based Protein Profiling (ABPP) of Oxidoreductases

Abstract

Over the last two decades, activity-based protein profiling (ABPP) has been established as a tremendously useful proteomic tool for measuring the activity of proteins in their cellular context, annotating the function of uncharacterized proteins, and investigating the target profile of small-molecule inhibitors. Unlike hydrolases and other enzyme classes, which exhibit a characteristic nucleophilic residue, oxidoreductases have received much less attention in ABPP. In this minireview, the state of the art of ABPP of oxidoreductases is described and the scope and limitations of the existing approaches are discussed. It is noted that several ABPP probes have been described for various oxidases, but none so far for a reductase, which gives rise to opportunities for future research.

Keywords: chemical proteomics; covalent inhibitors; drug discovery; oxidoreductases; proteomics.

Scheme 1

Design of the mechanism‐based P450‐probe 1. The warhead is depicted in red.

Figure 1

Selected probes designed for the ABPP of human P450s. The warheads are depicted in red.

Figure 2

Probe 7 developed for profiling cytochrome P450 activities associated with pyrethroid insecticide interactions. The warhead is depicted in red.

Figure 3

Benzofuran‐derived Probe 8 for P450 labeling. The warhead is depicted in red.

Scheme 2

Design of the mechanism‐based MAO‐probe 9. The warhead is depicted in red.

Figure 4

MAO B‐selective dual purpose probe 12. The warhead for covalent attachment is depicted in red, the chromophore for live‐cell imaging in blue.

Scheme 3

Labeling of flavin‐dependent oxidases with general probe 13. The warhead is depicted in red.

Figure 5

AMO‐probe 18. The warhead for covalent attachment is depicted in red.

Figure 6

MPO‐probe 20. The warhead for covalent attachment is depicted in red.

Scheme 4

15‐LOX‐1 probe 21 and its biorthogonal labeling by oxidative Heck reaction. The warhead is depicted in red.

Scheme 5

ABPP probe 25 for the visualization of LSDAO. The warhead is depicted in red.

Scheme 6

Oxidation of retinal to all‐trans retinoic acid (27) via ALDH1 A1 and the ABPP‐probe LEI‐945 (30). The warhead is depicted in red.

Figure 7

Broad‐spectrum aldehyde dehydrogenase ABPP probes 32 and STA‐55 (33) with a latent warhead.

Figure 8

ABPP probes for 2‐OG oxygenases (34), LSD‐1 (35), and PLP‐dependent enzymes (36).