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

Fluorescent photoaffinity probes for mitotic protein kinase Aurora A.

Author information

1
Institute of Chemistry, University of Tartu, Ravila 14A, 50411 Tartu, Estonia. Electronic address: darja.lavogina@ut.ee.
2
Institute of Chemistry, University of Tartu, Ravila 14A, 50411 Tartu, Estonia.

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 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)

PMID:
26077498
DOI:
10.1016/j.bmcl.2015.05.060
[Indexed for MEDLINE]

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