Fluorescent photoaffinity probes for mitotic protein kinase Aurora A

Darja Lavogina; Katariina Kisand; Gerda Raidaru; Asko Uri

doi:10.1016/j.bmcl.2015.05.060

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.

Authors

Darja Lavogina¹, Katariina Kisand², Gerda Raidaru², Asko Uri²

Affiliations

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.

PMID: 26077498
DOI: 10.1016/j.bmcl.2015.05.060

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).

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

Substances

Fluorescent Dyes
Photoaffinity Labels
Aurora Kinase A