| Title: | An Untargeted Approach for Revealing Electrophilic Metabolites |
| Author(s): | Yu Y; Le HH; Curtis BJ; Wrobel CJJ; Zhang B; Maxwell DN; Pan JY; Schroeder FC; |
| Address: | "Boyce Thompson Institute and Department of Chemistry and Chemical Biology, Cornell University, Ithaca, New York 14853, United States" |
| DOI: | 10.1021/acschembio.0c00706 |
| ISSN/ISBN: | 1554-8937 (Electronic) 1554-8929 (Print) 1554-8929 (Linking) |
| Abstract: | "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" |
| Keywords: | "Animals Aspergillus/metabolism *Biosynthetic Pathways Caenorhabditis elegans/metabolism Cell Line Chromatography, Liquid/methods Humans Hydroxylamine/*metabolism Isotope Labeling/methods Mass Spectrometry/methods Metabolome Metabolomics/methods;" |
| Notes: | "MedlineYu, Yan Le, Henry H Curtis, Brian J Wrobel, Chester J J Zhang, Bingsen Maxwell, Danielle N Pan, Judy Y Schroeder, Frank C eng R35 GM131877/GM/NIGMS NIH HHS/ T32 GM008500/GM/NIGMS NIH HHS/ HHMI/Howard Hughes Medical Institute/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't 2020/10/20 ACS Chem Biol. 2020 Nov 20; 15(11):3030-3037. doi: 10.1021/acschembio.0c00706. Epub 2020 Oct 19" |
