Chemoselective probes for metabolite enrichment and profiling. Carlson, E. E & Cravatt, B. F Nat Methods, 4(5):429–435, 2007. doi abstract bibtex 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.
@Article{carlson07chemoselective,
author = {Erin E Carlson and Benjamin F Cravatt},
title = {Chemoselective probes for metabolite enrichment and profiling.},
journal = {Nat Methods},
year = {2007},
volume = {4},
number = {5},
pages = {429--435},
abstract = {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.},
comment = {Understanding the metabolic and signaling networks that regulate health and disease is a principal goal of post-genomic research. To fully understand the composition and function of biochemical net- works, analysis of the small-molecule complement of cells and tissues, commonly referred to as the metabolome, is essential efforts to fully inventory the metabo- lome have been hindered by a unique set of challenges metabolites share no direct link to the genetic code, instead being synthesized by the concerted action of complex networks of enzymes metabolites constitute a structurally diverse collection of molecules that vary widely in physicochemical properties, including polarity, hydrophobicity, mass, chemical stability and relative abundance present estimates suggest that most eukaryotic organisms possess 4,000-20,000 distinct metabolites},
doi = {10.1038/nmeth1038},
keywords = {Acetylcysteine, pharmacology; Acrylic Resins; Antioxidants, pharmacology; Breast Neoplasms; Chromatography, Liquid; Humans; Polyethylene Glycols; Proteomics; Sulfhydryl Compounds; Tandem Mass Spectrometry; Trypsin; Tumor Cells, Cultured},
optmonth = may,
owner = {fhufsky},
pmid = {17417646},
timestamp = {2011.06.28},
}
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