Chemoselective probes for metabolite enrichment and profiling.
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.
Carlson
Erin E
EE
The Skaggs Institute for Chemical Biology, and Department of Cell Biology, The Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla, California 92037, USA.
Cravatt
Benjamin F
BF
D000111:Acetylcysteine / Q000494:pharmacology*; D000180:Acrylic Resins / Q000737:chemistry*; D000975:Antioxidants / Q000494:pharmacology*; D001943:Breast Neoplasms / Q000378:metabolism*; D002853:Chromatography, Liquid; D006801:Humans; D011092:Polyethylene Glycols / Q000737:chemistry*; D040901:Proteomics / Q000379:methods*; D013438:Sulfhydryl Compounds / Q000378:metabolism*; D053719:Tandem Mass Spectrometry; D014357:Trypsin / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000180:Acrylic Resins; D000975:Antioxidants; D013438:Sulfhydryl Compounds; C520358:poly(acryloyl-bis(aminopropyl)polyethylene glycol); D011092:Polyethylene Glycols; D014357:Trypsin; D000111:Acetylcysteine