Proteomic profiling of mechanistically distinct enzyme classes using a common chemotype

• Published in: Nature biotechnology Vol. 20; no. 8; pp. 805 - 809
• Main Authors: Cravatt, Benjamin F, Adam, Gregory C, Sorensen, Erik J
• Format: Journal Article
• Language: English
• Published: New York, NY Nature 01.08.2002; Nature Publishing Group
• Subjects: Affinity Labels - chemistry; Affinity Labels - metabolism; Analytical, structural and metabolic biochemistry; Animals; Binding Sites; Biological and medical sciences; Biotechnology; Breast Neoplasms - enzymology; Combinatorial Chemistry Techniques; COS Cells; Enzymes; Enzymes - chemistry; Enzymes - classification; Enzymes - genetics; Enzymes - metabolism; Enzymes and enzyme inhibitors; Fluorescent Dyes - chemistry; Fluorescent Dyes - metabolism; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; Glutathione Transferase - classification; Glutathione Transferase - metabolism; Health. Pharmaceutical industry; Humans; Immunoblotting; Industrial applications and implications. Economical aspects; Labeling; Mice; Miscellaneous; Molecular Probe Techniques; Molecular Structure; Probes; Protein expression; Proteins; Proteomics; Proteomics - methods; Sulfonates; Tumor Cells, Cultured; Screening; Molecular probe; Enzymatic activity; Enzyme; Affinity labelling; Proteomics
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/nbt714

Proteomics research requires methods to characterize the expression and function of proteins in complex mixtures. Toward this end, chemical probes that incorporate known affinity labeling agents have facilitated the activity-based profiling of certain enzyme families. To accelerate the discovery of proteomics probes for enzyme classes lacking cognate affinity labels, we describe here a combinatorial strategy. Members of a probe library bearing a sulfonate ester chemotype were screened against complex proteomes for activity-dependent protein reactivity, resulting in the labeling of at least six mechanistically distinct enzyme classes. Surprisingly, none of these enzymes represented targets of previously described proteomics probes. The sulfonate library was used to identify an omega-class glutathione S-transferase whose activity was upregulated in invasive human breast cancer lines. These results indicate that activity-based probes compatible with whole-proteome analysis can be developed for numerous enzyme classes and applied to identify enzymes associated with discrete pathological states.