Bioorthogonal oncometabolite ligation

Methods Enzymol. 2019:622:431-448. doi: 10.1016/bs.mie.2019.02.037. Epub 2019 Mar 14.


Dysregulated cellular metabolism is an emerging hallmark of cancer. Improved methods to profile aberrant metabolic activity thus have substantial applications as tools for diagnosis and understanding the biology of malignant tumors. Here we describe the utilization of a bioorthogonal ligation to fluorescently detect the TCA cycle oncometabolite fumarate. This method enables the facile measurement of fumarate hydratase activity in cell and tissue samples, and can be used to detect disruptions in metabolism that underlie the genetic cancer syndrome hereditary leiomyomatosis and renal cell cancer (HLRCC). The current method has substantial utility for sensitive fumarate hydratase activity profiling, and also provides a foundation for future applications in diagnostic detection and imaging of cancer metabolism.

Keywords: Bioorthogonal; Bioorthogonal oncometabolite ligation: HLRCC; Click chemistry; Cycloaddition; Fluorogenic; Fumarate; Fumarate hydratase; Hereditary cancer; Profiling; Sensor; Tetrazole.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • Citric Acid Cycle*
  • Click Chemistry / methods
  • Cycloaddition Reaction
  • Enzyme Assays / methods
  • Female
  • Fluorescent Dyes / analysis
  • Fluorescent Dyes / chemical synthesis
  • Fluorescent Dyes / metabolism
  • Fluorometry / methods
  • Fumarate Hydratase / metabolism*
  • Fumarates / analysis
  • Fumarates / metabolism*
  • Humans
  • Leiomyomatosis / metabolism*
  • Neoplastic Syndromes, Hereditary / metabolism*
  • Skin Neoplasms / metabolism*
  • Uterine Neoplasms / metabolism*


  • Fluorescent Dyes
  • Fumarates
  • Fumarate Hydratase

Supplementary concepts

  • Hereditary leiomyomatosis and renal cell cancer