title: Can we see PIP(3) and hydrogen peroxide with a single probe?
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.
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russia.
authors.forename: Natalie M
authors.initials: NM
authors.lastname: Mishina
authors.forename: Ivan
authors.initials: I
authors.lastname: Bogeski
authors.forename: Dmitry A
authors.initials: DA
authors.lastname: Bolotin
authors.forename: Markus
authors.initials: M
authors.lastname: Hoth
authors.forename: Barbara A
authors.initials: BA
authors.lastname: Niemeyer
authors.forename: Carsten
authors.initials: C
authors.lastname: Schultz
authors.forename: Eleva V
authors.initials: EV
authors.lastname: Zagaynova
authors.forename: Sergey
authors.initials: S
authors.lastname: Lukyanov
authors.forename: Vsevolod V
authors.initials: VV
authors.lastname: Belousov
mesh_terms: D000077329:Agammaglobulinaemia Tyrosine Kinase; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism*; D008164:Luminescent Proteins / Q000737:chemistry*; D051379:Mice; D008856:Microscopy, Fluorescence; D041681:NIH 3T3 Cells; D010084:Oxidation-Reduction; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism*; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology / Q000502:physiology; D017434:Protein Structure, Tertiary; D011505:Protein-Tyrosine Kinases / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000737:chemistry*; D015398:Signal Transduction; D013050:Spectrometry, Fluorescence; D006377:T-Lymphocytes, Helper-Inducer / Q000201:enzymology / Q000378:metabolism; D059008:Time-Lapse Imaging
chemical_list: D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D018129:Phosphatidylinositol Phosphates; D010982:Platelet-Derived Growth Factor; D011993:Recombinant Fusion Proteins; C060974:phosphatidylinositol 3,4,5-triphosphate; C054562:yellow fluorescent protein, Bacteria; D006861:Hydrogen Peroxide; D019869:Phosphatidylinositol 3-Kinases; D011505:Protein-Tyrosine Kinases; D000077329:Agammaglobulinaemia Tyrosine Kinase