4.5 Article

GDP-Capture Compound - A novel tool for the profiling of GTPases in pro- and eukaryotes by capture compound mass spectrometry (CCMS)

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JOURNAL OF PROTEOMICS
卷 73, 期 4, 页码 815-819

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ELSEVIER SCIENCE BV
DOI: 10.1016/j.jprot.2009.12.002

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Functional proteomics; Capture Compound Mass; Spectrometry; GTP; GTPases; Hek293; LC-MS/MS

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The functional isolation of proteome subsets based on small molecule-protein interactions is an increasingly popular and promising field in functional proteomics. Entire protein families may be profiled on the basis of their common interaction with a metabolite or small molecule inhibitor. This is enabled by novel multifunctional small molecule probes. One platform approach in this field are Capture Compounds that contain a small molecule of interest to bind target proteins, a photo-activatable reactivity function to covalently trap bound proteins, and a sorting function to isolate Capture Compound-protein conjugates from complex biological samples for direct trypsinisation and protein identification by liquid chromatography/mass spectrometry (CCMS). We here present the synthesis and application of a novel GDP-Capture Compound for the functional enrichment of GTPases, a pivotal protein family that exerts key functions in signal transduction. We present data from CCMS experiments on two biological lysates from Escherichia coli and from human-derived Hek293 cells. The GDP-Capture Compound robustly captures a wide range of different GTPases from both systems and will be a valuable tool for the proteomic profiling of this important protein family. (C) 2009 Elsevier B.V. All rights reserved.

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