Quantification of Cell Signaling Networks Using Kinase Activity Chemosensors

Authors

Jon R. Beck, University of Nebraska - Lincoln
Edward N. Harris, University of Nebraska - LincolnFollow
Cliff I. Stains, University of Nebraska-LincolnFollow

Date of this Version

2017

Citation

Published as Chapter 4 in Aik-Choon Tan and Paul H. Huang, eds., Kinase Signaling Networks, Methods in Molecular Biology, vol. 1636 (Springer, 2017), pp 61-70.

doi 10.1007/978-1-4939-7154-1_4,

PMID: 287304472

Comments

Copyright © 2017 Springer Science+Business Media LLC.

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.