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Activity-based protein profiling for the functional annotation of enzymes

Nature Methods volume 4, pages 822–827 (2007)Cite this article

Abstract

Activity-based protein profiling (ABPP), the use of active site-directed chemical probes to monitor enzyme function in complex biological systems, is emerging as a powerful post-genomic technology. ABPP probes have been developed for several enzyme classes and have been used to inventory enzyme activities en masse for a range of (patho) physiological processes. By presenting specific examples, we show here that ABPP provides researchers with a distinctive set of chemical tools to embark on the assignment of functions to many of the uncharacterized enzymes that populate eukaryotic and prokaryotic proteomes.

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Figure 1
Figure 2: Schematic of an MS experiment.
Figure 3: Examples of enzymes assigned to specific mechanistic classes by ABPP.
Figure 4: Substrate mimicry of an ABPP probe.
Figure 5: Inhibitor screening by competitive ABPP.
Figure 6: Multidimensional profiling strategy for the annotation of the cancer-related enzyme KIAA1363.

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Acknowledgements

We thank members of the Cravatt lab for careful reading of this manuscript and gratefully acknowledge the support of the US National Institutes of Health (CA087660, CA118696), the Helen L. Dorris Child and Adolescent Neuropsychiatric Disorder Institute and the Skaggs Institute for Chemical Biology.

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  1. The Skaggs Institute for Chemical Biology and Department of Chemical Physiology, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, 92037, California, USA

    Katherine T Barglow & Benjamin F Cravatt

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Barglow, K., Cravatt, B. Activity-based protein profiling for the functional annotation of enzymes. Nat Methods 4, 822–827 (2007). https://doi.org/10.1038/nmeth1092

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