Comparison of the efficiencies of two TR-FRET methods to detect in vitro natural and synthesized inhibitors of the Raf/MEK/ERK signaling pathway

mparison of the efficiencies of two TR-FRET methods to detect in vitro natural and synthesized inhibitors of the Raf/MEK/ERK signaling pathway Original Research (3859) Total Article Views Authors: Fabienne Saab, Sylvain Routier, Jean-Yves Mérour, et al Published Date June 2010 Volume 2010:1 Pages 81 - 98 DOI: http://dx.doi.org/10.2147/IJHTS.S9157 Fabienne Saab1,2, Sylvain Routier2, Jean-Yves Mérour2, Valérie Bénéteau2, Fran oise Schoentgen1,3 1Centre de Biophysique Moléculaire UPR4301 CNRS, affiliated to Orleans University and to INSERM, France; 2Institut de Chimie Organique et Analytique UMR6005 CNRS, Université d’Orléans, France; 3IMPMC UMR 7590 CNRS, Université Pierre et Marie Curie, France Abstract: Numerous types of cancer operate through the deregulation of the Raf/MEK/ERK pathway. It is therefore of importance to design and synthesize inhibitors of this pathway. Consequently, we have developed several tests to measure in vitro the effect of inhibitors on the activity of the complete cascade Raf-1/MEK/ERK and also on the activities of Raf-1, MEK, and ERK individually. We present here the results obtained with two time-resolved fluorescence resonance energy transfer (TR-FRET) methods by comparison with a classical radioactivity method and experimental data found in literature. The capability of a series of optimized assays to detect different types of inhibitors is evaluated and discussed.