Comparison of LanthaScreen Eu kinase binding assay and surface plasmon resonance method in elucidating the binding kinetics of focal adhesion kinase inhibitors.
An understanding of the dynamics of drug-target interactions is important in the drug discovery process. Information related to the binding kinetics of a drug toward its target or off-target aids in determining the efficacy or toxicity of a drug. Biophysical techniques such as surface plasmon resonance (SPR) have been available for over 20 years, but have been predominantly utilized to characterize protein-protein interactions. With improvements in instrument sensitivity and data analysis software, interactions between proteins (such as kinases) and small molecules have been successfully evaluated. More recently, the LanthaScreen Eu kinase binding assay for characterizing kinase inhibitors has been described. This assay monitors displacement of an Alexa Fluor 647-labeled tracer from the ATP-binding site of an epitope-tagged kinase by a test compound. Such behavior results in a decrease in time-resolved fluorescence energy transfer signal. In this report, a side-by-side comparison of the LanthaScreen Eu kinase binding assay and the SPR method was performed using inhibitors of focal adhesion kinase. The two methods yielded comparable results and identified compounds with time-dependent inhibition and relatively slow dissociation.
Worldwide Discovery Research, Cephalon, Inc., West Chester, Pennsylvania, USA.
Jennifer L
JL
Mason
Chrysanthe
C
Spais
Jean
J
Husten
Eric
E
Prouty
Mark S
MS
Albom
Sheryl L
SL
Meyer
Mark A
MA
Ator
Thelma S
TS
Angeles
D051417:Focal Adhesion Kinase 1 / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D006801:Humans; D011485:Protein Binding / Q000187:drug effects / Q000502:physiology; D047428:Protein Kinase Inhibitors / Q000378:metabolism* / Q000493:pharmacokinetics; D019311:Staurosporine / Q000378:metabolism / Q000493:pharmacokinetics; D020349:Surface Plasmon Resonance / Q000379:methods*
D047428:Protein Kinase Inhibitors; D051417:Focal Adhesion Kinase 1; C496050:PTK2 protein, human; D019311:Staurosporine