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Research

Assessing Enzyme Activities Using Stable Isotope Labeling and Mass Spectrometry ^*,S

Patrick A. Everley^,, Carlos A. Gartner, Wilhelm Haas, Alan Saghatelian^¶, Joshua E. Elias, Benjamin F. Cravatt^¶, Bruce R. Zetter and Steven P. Gygi^,||

From the Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, Program in Vascular Biology, Department of Surgery, Children's Hospital, Boston, Massachusetts 02115, and ^¶ The Skaggs Institute for Chemical Biology and Department of Cell Biology, The Scripps Research Institute, La Jolla, California 92037

Activity-based protein profiling has emerged as a valuable technology for labeling, enriching, and assessing protein activities from complex mixtures. This is primarily accomplished via a two-step identification and quantification process. Here we show a highly quantitative and streamlined method, termed catch-and-release activity profiling of enzymes (CAPE), which reduces this procedure to a single step. Furthermore the CAPE approach has the ability to detect small quantitative changes that may have been missed by alternative mass spectrometry-based techniques.