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Chemoselective probes for metabolite enrichment and profiling

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Overview

authors

  • Carlson, E. E.
  • Cravatt, Benjamin

publication date

  • May 2007

journal

  • Nature Methods  Journal

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.

subject areas

  • Acetylcysteine
  • Acrylic Resins
  • Antioxidants
  • Breast Neoplasms
  • Chromatography, Liquid
  • Humans
  • Polyethylene Glycols
  • Proteomics
  • Sulfhydryl Compounds
  • Tandem Mass Spectrometry
  • Trypsin
  • Tumor Cells, Cultured
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Identity

International Standard Serial Number (ISSN)

  • 1548-7091

Digital Object Identifier (DOI)

  • 10.1038/nmeth1038

PubMed ID

  • 17417646
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Additional Document Info

start page

  • 429

end page

  • 435

volume

  • 4

issue

  • 5

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