Modular activatable bioorthogonal reagents

Methods Enzymol. 2019:622:153-182. doi: 10.1016/bs.mie.2019.02.007. Epub 2019 Mar 6.


The bioorthogonal reaction toolbox contains approximately two-dozen unique chemistries that permit selective tagging and probing of biomolecules. Over the past two decades, significant effort has been devoted to optimizing and discovering bioorthogonal reagents that are faster, fluorogenic, and orthogonal to the already existing bioorthogonal repertoire. Conversely, efforts to explore bioorthogonal reagents whose reactivity can be controlled in space and/or time are limited. The "activatable" bioorthogonal reagents that do exist are often unimodal, meaning that their reagent's activation method cannot be easily modified to enable activation with red-shifted wavelengths, enzymes, or metabolic-byproducts and ions like H2O2 or Fe3+. Here, we summarize the available activatable bioorthogonal reagents with a focus on our recent addition: modular caged cyclopropenes. We designed caged cyclopropenes to be unreactive to their bioorthogonal partner until they are activated through the removal of the cage by light, an enzyme, or another reaction partner. To accomplish this, their structure includes a nitrogen atom at the cyclopropene C3 position that is decorated with the desired caging group through a carbamate linkage. This 3-N cyclopropene system can allow control of cyclopropene reactivity using a multitude of already available photo- and enzyme-caging groups. Additionally, this cyclopropene scaffold can enable metabolic-byproduct or ion activation of bioorthogonal reactions.

Keywords: 3-N cyclopropenes; Bioorthogonal reactions; Caged cyclopropenes; Enzyme cage; Enzyme-activatable bioorthogonal reactions; Light-activatable bioorthogonal reactions; Photocage; Spatiotemporal control of bioorthogonal reactions.

MeSH terms

  • Animals
  • Biocatalysis
  • Carbamates / chemical synthesis
  • Carbamates / chemistry
  • Chemistry Techniques, Synthetic / methods
  • Chromatography, High Pressure Liquid / methods
  • Cyclopropanes / chemical synthesis
  • Cyclopropanes / chemistry*
  • Humans
  • Hydrogen Peroxide / chemistry
  • Indicators and Reagents
  • Light
  • Nitrogen / chemistry


  • Carbamates
  • Cyclopropanes
  • Indicators and Reagents
  • Hydrogen Peroxide
  • Nitrogen