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A Rapidly Reversible Chemical Dimerizer System to Study Lipid Signaling in Living Cells
Author(s) -
Feng Suihan,
Laketa Vibor,
Stein Frank,
Rutkowska Anna,
MacNamara Aidan,
Depner Sofia,
Klingmüller Ursula,
SaezRodriguez Julio,
Schultz Carsten
Publication year - 2014
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201402294
Subject(s) - phosphatidylinositol , effector , chemistry , confocal microscopy , enzyme , biophysics , microbiology and biotechnology , fluorescence microscope , kinase , biochemistry , fluorescence , biology , physics , quantum mechanics
Chemical dimerizers are powerful tools for non‐invasive manipulation of enzyme activities in intact cells. Here we introduce the first rapidly reversible small‐molecule‐based dimerization system and demonstrate a sufficiently fast switch‐off to determine kinetics of lipid metabolizing enzymes in living cells. We applied this new method to induce and stop phosphatidylinositol 3‐kinase (PI3K) activity, allowing us to quantitatively measure the turnover of phosphatidylinositol 3,4,5‐trisphosphate (PIP 3 ) and its downstream effectors by confocal fluorescence microscopy as well as standard biochemical methods.