Fluorescent photoaffinity probes for mitotic protein kinase Aurora A. Issue 16 (15th August 2015)

• Record Type: Journal Article
• Authors: Lavogina, Darja; Kisand, Katariina; Raidaru, Gerda; Uri, Asko
• Is Part Of: Bioorganic & medicinal chemistry letters. Volume 25:Issue 16(2015)
• Page Start: 3290
• Page End: 3294
• Publication Date: 2015-08-15
• Subjects: Protein kinase -- Aurora A -- Photoaffinity labelling -- Fluorescent probe -- Covalent irreversible inhibitor -- VX689 (MK-5108); Bioorganic chemistry -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1016/j.bmcl.2015.05.060
• Languages: English
• ISSNs: 0960-894X

Abstract

Graphical abstract: Abstract: 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 K D 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.