Quantification of cell signaling networks using kinase activity chemosensors

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages61-70
Number of pages10
DOIs
StatePublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1636
ISSN (Print)1064-3745

Keywords

  • Cell signaling
  • Fluorescence-based biosensor
  • Kinase activity assay
  • Nonalcoholic fatty liver disease
  • Phosphorylation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

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