Site-specific incorporation of quadricyclane into a protein and photocleavage of the quadricyclane ligation adduct

Bioorg Med Chem. 2018 Oct 15;26(19):5280-5290. doi: 10.1016/j.bmc.2018.04.009. Epub 2018 Apr 4.

Abstract

The quadricyclane (QC) ligation is a bioorthogonal reaction between a quadricyclane moiety and a nickel bis(dithiolene) derivative. Here we show that a QC amino acid can be incorporated into a protein site-specifically using the pyrrolysine-based genetic code expansion platform, and subsequently used for ligation chemistry. Additionally, we exploited the photolability of the QC ligation product to render the adduct cleavable with a handheld UV lamp. We further developed a protein purification method that involves QC ligation of biotin to a protein of interest, capture on streptavidin resin, and finally release using only UV light. The QC ligation thus brings novel chemical manipulations to the realm of bioorthogonal chemistry.

Keywords: Bioorthogonal; Photocleavage; Photolysis; Protein purification; Pyrrolysine; Quadricyclane; Unnatural amino acid.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Biotin / chemistry*
  • Biotin / metabolism
  • Bridged-Ring Compounds / chemistry*
  • Coordination Complexes / chemistry
  • Lysine / analogs & derivatives
  • Lysine / chemistry
  • Nickel / chemistry
  • Photolysis / radiation effects
  • Streptavidin / chemistry
  • Streptavidin / metabolism
  • Ultraviolet Rays*

Substances

  • Bridged-Ring Compounds
  • Coordination Complexes
  • Biotin
  • Nickel
  • quadricyclane
  • Streptavidin
  • pyrrolysine
  • Lysine