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Quantification Of Cell Signaling Networks Using Kinase Activity Chemosensors, Jon R. Beck, Edward N. Harris, Cliff I. Stains
Quantification Of Cell Signaling Networks Using Kinase Activity Chemosensors, Jon R. Beck, Edward N. Harris, Cliff I. Stains
Department of Biochemistry: Faculty Publications
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 …