Quantification of cell signaling networks using kinase activity chemosensors

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Abstract

The ability to directly determine endogenous kinase activity in tissue homogenates provides valuable insights into signaling aberrations that underlie disease phenotypes. When activity data is collected across a panel of kinases, a unique “signaling fingerprint” is generated that allows for discrimination between diseased and normal tissue. Here we describe the use of peptide-based kinase activity sensors to fingerprint the signaling changes associated with disease states. This approach leverages the phosphorylation-sensitive sulfonamido-oxine (Sox) fluorophore to provide a direct readout of kinase enzymatic activity in unfractionated tissue homogenates from animal models or clinical samples. To demonstrate the application of this technology, we focus on a rat model of nonalcoholic fatty liver disease (NAFLD). Sox-based activity probes allow for the rapid and straightforward analysis of changes in kinase enzymatic activity associated with disease states, providing leads for further investigation using traditional biochemical approaches.

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Beck, J. R., Harris, E. N., & Stains, C. I. (2017). Quantification of cell signaling networks using kinase activity chemosensors. In Methods in Molecular Biology (Vol. 1636, pp. 61–70). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7154-1_4

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