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Chemical approaches for functionally probing the proteome.

Doron Greenbaum·Amos Baruch·Linda Hayrapetian·Zsuzsanna Darula·Alma Burlingame·Katlin F Medzihradszky·Matthew Bogyo
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Abstract

With the availability of complete genome sequences, emphasis has shifted toward the understanding of protein function. We have developed a functional proteomic methodology that makes use of chemically reactive fluorescent probes to profile and identify enzymes in complex mixtures by virtue of their catalytic activity. This methodology allows a comparison of changes in activity of multiple enzymes under a variety of conditions using a single two-dimensional separation. The probes can also be used to localize active enzymes in intact cells using fluorescence microscopy. Furthermore, the probes enable screens for selective small molecule inhibitors of each enzyme family member within crude lysates or intact cells. Ultimately, this technology allows the rapid identification of potential drug targets and small molecule lead compounds targeted to them.

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Affinity LabelsAnimalsBinding SitesCathepsinsEnzyme InhibitorsFluorescent DyesIndicators and ReagentsLiverMolecular Probe TechniquesMolecular StructurePeptide FragmentsPeptide HydrolasesPeptide LibraryProteomeRatsSerine EndopeptidasesSpectrometry, FluorescenceJournal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, P.H.S.

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Doron Greenbaum, Amos Baruch, Linda Hayrapetian, Zsuzsanna Darula, Alma Burlingame, Katlin F Medzihradszky, Matthew Bogyo. 2002. Chemical approaches for functionally probing the proteome.. https://doi.org/10.1074/mcp.t100003-mcp200

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