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Originally published In Press as doi:10.1074/mcp.M700057-MCP200 on July 11, 2007.
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Molecular & Cellular Proteomics 6:1771-1777, 2007.
© 2007 by The American Society for Biochemistry and Molecular Biology, Inc.

Research

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

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

From the {ddagger} 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.

|| To whom correspondence should be addressed. Tel.: 617-432-3155; Fax: 617-432-1144; E-mail: steven_gygi{at}hms.harvard.edu


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L. C. J. Gillet, K. Namoto, A. Ruchti, S. Hoving, D. Boesch, B. Inverardi, D. Mueller, M. Coulot, P. Schindler, P. Schweigler, et al.
In-cell Selectivity Profiling of Serine Protease Inhibitors by Activity-based Proteomics
Mol. Cell. Proteomics, July 1, 2008; 7(7): 1241 - 1253.
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Copyright © 2007 by the American Society for Biochemistry and Molecular Biology.