Bioorthogonal click chemistry to assay mu-opioid receptor palmitoylation using 15-hexadecynoic acid and immunoprecipitation

Anal Biochem. 2014 Apr 15:451:25-7. doi: 10.1016/j.ab.2014.01.008. Epub 2014 Jan 23.


We have developed a modification of bioorthogonal click chemistry to assay the palmitoylation of cellular proteins. This assay uses 15-hexadecynoic acid (15-HDYA) as a chemical probe in combination with protein immunoprecipitation using magnetic beads in order to detect S-palmitoylation of proteins of interest. Here we demonstrate the utility of this approach for the mu-opioid receptor (MOR), a G-protein-coupled receptor (GPCR) responsible for mediating the analgesic and addictive properties of most clinically relevant opioid agonist drugs. This technique provides a rapid, non-isotopic, and efficient method to assay the palmitoylation status of a variety of cellular proteins, including most GPCRs.

Keywords: Click chemistry; Immunoprecipitation; Mu-opioid receptor; Palmitoylation.

Publication types

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

MeSH terms

  • Antibodies / immunology
  • Click Chemistry
  • Fatty Acids, Unsaturated / chemistry*
  • HEK293 Cells
  • Humans
  • Immunoblotting*
  • Immunomagnetic Separation
  • Immunoprecipitation
  • Lipoylation
  • Receptors, Opioid, mu / analysis*
  • Receptors, Opioid, mu / genetics
  • Receptors, Opioid, mu / metabolism
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / isolation & purification


  • Antibodies
  • Fatty Acids, Unsaturated
  • Receptors, Opioid, mu
  • Recombinant Fusion Proteins
  • 15-hexadecynoic acid