Open AccessAn Untargeted Approach for Revealing Electrophilic Metabolites
Author(s) -
Yang Yu,
Henry Le,
Brian J Curtis,
Chester J J Wrobel,
Bingsen Zhang,
Danielle N Maxwell,
Judy Y Pan,
Frank C. Schroeder
Publication year - 2020
Publication title -
acs chemical biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.899
H-Index - 111
eISSN - 1554-8937
pISSN - 1554-8929
DOI - 10.1021/acschembio.0c00706
Subject(s) - thioesterase , electrophile , caenorhabditis elegans , chemistry , biochemistry , biosynthesis , catabolism , biology , metabolism , gene , catalysis
Reactive electrophilic intermediates such as coenzyme A esters play central roles in metabolism but are difficult to detect with conventional strategies. Here, we introduce hydroxylamine-based stable isotope labeling to convert reactive electrophilic intermediates into stable derivatives that are easily detectable via LC-MS. In the model system Caenorhabditis elegans , parallel treatment with 14 NH 2 OH and 15 NH 2 OH revealed >1000 labeled metabolites, e.g., derived from peptide, fatty acid, and ascaroside pheromone biosyntheses. Results from NH 2 OH treatment of a pheromone biosynthesis mutant, acox-1.1 , suggested upregulation of thioesterase activity, which was confirmed by gene expression analysis. The upregulated thioesterase contributes to the biosynthesis of a specific subset of ascarosides, determining the balance of dispersal and attractive signals. These results demonstrate the utility of NH 2 OH labeling for investigating complex biosynthetic networks. Initial results with Aspergillus and human cell lines indicate applicability toward uncovering reactive metabolomes in diverse living systems.