Can we see PIP(3) and hydrogen peroxide with a single probe?

Antioxid Redox Signal. 2012 Aug 1;17(3):505-12. doi: 10.1089/ars.2012.4574. Epub 2012 Apr 10.

Abstract

A genetically encoded sensor for parallel measurements of phosphatidylinositol 3-kinase activity and hydrogen peroxide (H(2)O(2)) levels (termed PIP-SHOW) was developed. Upon elevation of local phosphatidylinositol 3,4,5-trisphosphate (PIP(3)) concentration, the sensor translocates from the cytosol to the plasma membrane, while a ratiometric excitation change rapidly and simultaneously reports changes in the concentration of H(2)O(2). The dynamics of PIP(3) and H(2)O(2) generation were monitored in platelet-derived growth factor-stimulated fibroblasts and in T-lymphocytes after formation of an immunological synapse. We suggest that PIP-SHOW can serve as a prototype for many fluorescent sensors with combined readouts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agammaglobulinaemia Tyrosine Kinase
  • Animals
  • Bacterial Proteins / chemistry*
  • Fluorescent Dyes / chemistry*
  • HeLa Cells
  • Humans
  • Hydrogen Peroxide / metabolism*
  • Luminescent Proteins / chemistry*
  • Mice
  • Microscopy, Fluorescence
  • NIH 3T3 Cells
  • Oxidation-Reduction
  • Phosphatidylinositol 3-Kinases / metabolism
  • Phosphatidylinositol Phosphates / metabolism*
  • Platelet-Derived Growth Factor / pharmacology
  • Platelet-Derived Growth Factor / physiology
  • Protein Structure, Tertiary
  • Protein-Tyrosine Kinases / chemistry*
  • Recombinant Fusion Proteins / chemistry*
  • Signal Transduction
  • Spectrometry, Fluorescence
  • T-Lymphocytes, Helper-Inducer / enzymology
  • T-Lymphocytes, Helper-Inducer / metabolism
  • Time-Lapse Imaging

Substances

  • Bacterial Proteins
  • Fluorescent Dyes
  • Luminescent Proteins
  • Phosphatidylinositol Phosphates
  • Platelet-Derived Growth Factor
  • Recombinant Fusion Proteins
  • phosphatidylinositol 3,4,5-triphosphate
  • yellow fluorescent protein, Bacteria
  • Hydrogen Peroxide
  • Phosphatidylinositol 3-Kinases
  • Protein-Tyrosine Kinases
  • Agammaglobulinaemia Tyrosine Kinase