Assessing cellular protein phosphorylation: high throughput drug discovery technologies.

Russello, S.V.
Journal   Assay and Drug Dev Technol.
Species  
Analytes Measured  
Matrix Tested  
Year   2004
Volume   2
Page Numbers   225-35
Application   Phosphoproteins
Abstract
Changes in protein phosphorylation mediate much of cellular physiology. Perturbations in the activity of the kinases that catalyze these reactions underlie numerous human pathologies, including metabolic and inflammatory disorders and most notably, cancer. HTS techniques that determine the activity of protein kinases in vitro are useful in the development of small molecule kinase inhibitors, but do not address underlying mechanistic concerns or efficient in vivo targeting. Observing protein phosphorylation in cell lysates and fixed cells in a high throughput manner is fundamental to understanding the mechanism of action of lead molecules and whether they target signaling pathways of interest. Herein we discuss several higher throughput techniques to study cellular protein kinase signal transduction and the strategies for implementation in kinase drug discovery.

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