Interrogating signaling nodes involved in cellular transformations using kinase activity probes

Clifford I Stains, Nathan C. Tedford, Traci C. Walkup, Elvedin Luković, Brenda N. Goguen, Linda G. Griffith, Douglas A. Lauffenburger, Barbara Imperiali

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Protein kinases catalyze protein phosphorylation and thereby control the flow of information through signaling cascades. Currently available methods for concomitant assessment of the enzymatic activities of multiple kinases in complex biological samples rely on indirect proxies for enzymatic activity, such as posttranslational modifications to protein kinases. Our laboratories have recently described a method for directly quantifying the enzymatic activity of kinases in unfractionated cell lysates using substrates containing a phosphorylation-sensitive unnatural amino acid termed CSox, which can be monitored using fluorescence. Here, we demonstrate the utility of this method using a probe set encompassing p38α, MK2, ERK1/2, Akt, and PKA. This panel of chemosensors provides activity measurements of individual kinases in a model of skeletal muscle differentiation and can be readily used to generate individualized kinase activity profiles for tissue samples from clinical cancer patients.

Original languageEnglish (US)
Pages (from-to)210-217
Number of pages8
JournalChemistry and Biology
Volume19
Issue number2
DOIs
StatePublished - Feb 24 2012

Fingerprint

Phosphotransferases
Phosphorylation
Protein Kinases
Proxy
Post Translational Protein Processing
Muscle
Skeletal Muscle
Fluorescence
Tissue
Amino Acids
Substrates
Neoplasms
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Drug Discovery
  • Molecular Biology
  • Clinical Biochemistry
  • Molecular Medicine
  • Pharmacology

Cite this

Stains, C. I., Tedford, N. C., Walkup, T. C., Luković, E., Goguen, B. N., Griffith, L. G., ... Imperiali, B. (2012). Interrogating signaling nodes involved in cellular transformations using kinase activity probes. Chemistry and Biology, 19(2), 210-217. https://doi.org/10.1016/j.chembiol.2011.11.012

Interrogating signaling nodes involved in cellular transformations using kinase activity probes. / Stains, Clifford I; Tedford, Nathan C.; Walkup, Traci C.; Luković, Elvedin; Goguen, Brenda N.; Griffith, Linda G.; Lauffenburger, Douglas A.; Imperiali, Barbara.

In: Chemistry and Biology, Vol. 19, No. 2, 24.02.2012, p. 210-217.

Research output: Contribution to journalArticle

Stains, CI, Tedford, NC, Walkup, TC, Luković, E, Goguen, BN, Griffith, LG, Lauffenburger, DA & Imperiali, B 2012, 'Interrogating signaling nodes involved in cellular transformations using kinase activity probes', Chemistry and Biology, vol. 19, no. 2, pp. 210-217. https://doi.org/10.1016/j.chembiol.2011.11.012
Stains, Clifford I ; Tedford, Nathan C. ; Walkup, Traci C. ; Luković, Elvedin ; Goguen, Brenda N. ; Griffith, Linda G. ; Lauffenburger, Douglas A. ; Imperiali, Barbara. / Interrogating signaling nodes involved in cellular transformations using kinase activity probes. In: Chemistry and Biology. 2012 ; Vol. 19, No. 2. pp. 210-217.
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