Univ Rennes, CNRS, IGDR (Genetics and Development Institute of Rennes), UMR 6290, F-35000 Rennes, France

1 ARTICLES PUBLISHED IN JoVE

Biology

Real-Time Monitoring of Aurora kinase A Activation using Conformational FRET Biosensors in Live Cells

Giulia Bertolin¹, Gilles Le Marchand¹, Marc Tramier¹

¹Univ Rennes, CNRS, IGDR (Genetics and Development Institute of Rennes), UMR 6290, F-35000 Rennes, France

The activation of the multifunctional Ser/Thr kinase AURKA is hallmarked by its autophosphorylation on Thr288. Fluorescent probes relying on FRET can discriminate between its inactive and activated states. Here, we illustrate some strategies for probe engineering, together with a rapid FRET protocol to follow the kinase activation throughout mitosis.