Design and synthesis of fluorescent activity probes for protein phosphatases

Garrett R. Casey, Jon R. Beck, Cliff I. Stains

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Citation (Scopus)

Abstract

Protein phosphatases act in concert with protein kinases to regulate and maintain the phosphoproteome. However, the catalog of chemical tools to directly monitor the enzymatic activity of phosphatases has lagged behind their kinase counterparts. In this chapter, we provide protocols for repurposing the phosphorylation-sensitive sulfonamido-oxine fluorophore known as Sox to afford direct activity probes for phosphatases. With validated activity probes in-hand, inhibitor screens can be conducted with recombinant enzyme and the role of phosphatases in cell signaling can be investigated in unfractionated cell lysates.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
EditorsArun K. Shukla
PublisherAcademic Press Inc.
Pages29-53
Number of pages25
ISBN (Print)9780128181195
DOIs
StatePublished - 2019

Publication series

NameMethods in Enzymology
Volume622
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Fingerprint

Phosphoprotein Phosphatases
Fluorescent Dyes
Phosphoric Monoester Hydrolases
Cell signaling
Oxyquinoline
Phosphorylation
Fluorophores
Protein Kinases
Phosphotransferases
Hand
Enzymes

Keywords

  • Biosensor
  • Cell signaling
  • Fluorescent probes
  • Phosphatase
  • Sensor

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Casey, G. R., Beck, J. R., & Stains, C. I. (2019). Design and synthesis of fluorescent activity probes for protein phosphatases. In A. K. Shukla (Ed.), Methods in Enzymology (pp. 29-53). (Methods in Enzymology; Vol. 622). Academic Press Inc.. https://doi.org/10.1016/bs.mie.2019.02.002

Design and synthesis of fluorescent activity probes for protein phosphatases. / Casey, Garrett R.; Beck, Jon R.; Stains, Cliff I.

Methods in Enzymology. ed. / Arun K. Shukla. Academic Press Inc., 2019. p. 29-53 (Methods in Enzymology; Vol. 622).

Research output: Chapter in Book/Report/Conference proceedingChapter

Casey, GR, Beck, JR & Stains, CI 2019, Design and synthesis of fluorescent activity probes for protein phosphatases. in AK Shukla (ed.), Methods in Enzymology. Methods in Enzymology, vol. 622, Academic Press Inc., pp. 29-53. https://doi.org/10.1016/bs.mie.2019.02.002
Casey GR, Beck JR, Stains CI. Design and synthesis of fluorescent activity probes for protein phosphatases. In Shukla AK, editor, Methods in Enzymology. Academic Press Inc. 2019. p. 29-53. (Methods in Enzymology). https://doi.org/10.1016/bs.mie.2019.02.002
Casey, Garrett R. ; Beck, Jon R. ; Stains, Cliff I. / Design and synthesis of fluorescent activity probes for protein phosphatases. Methods in Enzymology. editor / Arun K. Shukla. Academic Press Inc., 2019. pp. 29-53 (Methods in Enzymology).
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