Fluorescent photoaffinity probes for mitotic protein kinase Aurora A

Bioorg Med Chem Lett. 2015 Aug 15;25(16):3290-4. doi: 10.1016/j.bmcl.2015.05.060. Epub 2015 May 30.


We combined the advantages of the selective inhibitor VX689, the bisubstrate-analogue conjugate approach, and photoreactive amino acids to develop 8 photoaffinity probes for Aurora A. The most efficient compounds possessed one-digit nanomolar KD values in the equilibrium binding assay, inhibited Aurora A at elevated concentrations of ATP in the phosphorylation assay in the presence of TPX2, and formed covalent complexes with the recombinant kinase or Aurora A in HeLa cells upon UV-irradiation. The recognition of the correct target by the probes during formation of the covalent complex in the biochemical assay and in situ was demonstrated by competition experiments using the non-labelled inhibitors VX689 and MLN8237.

Keywords: Aurora A; Covalent irreversible inhibitor; Fluorescent probe; Photoaffinity labelling; Protein kinase; VX689 (MK-5108).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aurora Kinase A / metabolism*
  • Biological Assay
  • Enzyme Assays / methods*
  • Fluorescent Dyes / chemical synthesis*
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / metabolism*
  • HeLa Cells
  • Humans
  • Molecular Structure
  • Photoaffinity Labels / chemical synthesis
  • Photoaffinity Labels / chemistry


  • Fluorescent Dyes
  • Photoaffinity Labels
  • Aurora Kinase A