Nature Methods
- 4, 429 - 435 (2007)
Published online: 8 April 2007; | doi:10.1038/nmeth1038
Chemoselective probes for metabolite enrichment and profilingErin E Carlson & Benjamin F Cravatt
The Skaggs Institute for Chemical Biology, and Departments of Cell Biology and Chemistry, The Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla, California 92037, USA.
Correspondence should be addressed to Benjamin F Cravatt cravatt@scripps.edu Chemical probes that target classes of proteins based on shared functional properties have emerged as powerful tools for proteomics. The metabolome rivals, if not surpasses, the proteome in terms of size and complexity, suggesting that efforts to profile metabolites would also benefit from targeted technologies. Here we apply the principle of chemoselective probes to the metabolome, creating a general strategy to tag, enrich and profile large classes of small molecules from biological systems. Key to success was incorporation of a protease-cleavage step to release captured metabolites in a format compatible with liquid chromatography–mass spectrometry (LC-MS) analysis. This technology, termed metabolite enrichment by tagging and proteolytic release (METPR), is applicable to small molecules of any physicochemical class, including polar, labile and low-mass (<100 Da) compounds. We applied METPR to profile changes in the thiol metabolome of human cancer cells treated with the antioxidant N-acetyl-L-cysteine.
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