Preparation and properties of an improved photoaffinity ligand for the N-formyl peptide receptor

Biochim Biophys Acta. 1986 Jul 16;882(3):271-80. doi: 10.1016/0304-4165(86)90248-5.


A new superior photoaffinity ligand for the N-formyl peptide receptor was prepared by derivatization of N-formyl-Met-Leu-Phe-Lys with a commercially available heterobifunctional crosslinking agent. The product, N-formyl-Met-Leu-Phe-N epsilon-(2-(p-azidosalicylamido)ethyl-1,3'- dithiopropionyl)-Lys was readily synthesized and radiolabelled, and had increased specificity and stability as compared to previously used photoaffinity ligands. The ligand rapidly associated with the receptor with high affinity (Kd = 0.28 nM). Once bound, it was virtually non-dissociable (in the absence of photolysis) in an experimental time-frame (t1/2 (off) = 154 min). The covalent incorporation of the photoaffinity ligand into the receptor upon irradiation was complete within 5 min, minimizing cell damage, and the efficiency of covalent incorporation was approx. 40%. The derivative had enhanced biological activity, having an ED50 for superoxide anion production of 0.23 nM, 27-fold lower than its parent peptide. This derivative of the N-formyl peptide was useful for up to 3 months after radiolabelling, showing a progressive decline in specific activity during storage in the dark at 4 degrees C. The enhanced stability, reproducibility and solubility of the photoaffinity ligand is expected to aid in the purification of the N-formyl peptide receptor and will prove a useful tool for analysing receptor-mediated processes.

Publication types

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

MeSH terms

  • Affinity Labels / chemical synthesis*
  • Affinity Labels / metabolism
  • Azides / chemical synthesis*
  • Chromatography, High Pressure Liquid
  • Cross-Linking Reagents / metabolism
  • Humans
  • Kinetics
  • N-Formylmethionine Leucyl-Phenylalanine / analogs & derivatives*
  • N-Formylmethionine Leucyl-Phenylalanine / chemical synthesis
  • Neutrophils / metabolism
  • Photochemistry
  • Receptors, Formyl Peptide
  • Receptors, Immunologic / metabolism*
  • Substrate Specificity
  • Superoxides / metabolism
  • Time Factors


  • Affinity Labels
  • Azides
  • Cross-Linking Reagents
  • Receptors, Formyl Peptide
  • Receptors, Immunologic
  • N-formyl-methionylleucyl-phenylalanyl-N(epsilon)-(2-(4-azidosalicylamido)ethyl-1,3'-dithiopropionyl)lysine
  • Superoxides
  • N-Formylmethionine Leucyl-Phenylalanine