TY - JOUR
T1 - A rapidly reversible chemical dimerizer system to study lipid signaling in living cells
AU - Feng, Suihan
AU - Laketa, Vibor
AU - Stein, Frank
AU - Rutkowska, Anna
AU - MacNamara, Aidan
AU - Depner, Sofia
AU - Klingmüller, Ursula
AU - Saez-Rodriguez, Julio
AU - Schultz, Carsten
PY - 2014/6/23
Y1 - 2014/6/23
N2 - 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. Little helper: Chemical dimerizers are powerful tools for manipulation of enzyme activities in intact cells. The first rapidly reversible chemical dimerization system is introduced, which permits to determine kinetics of lipid metabolizing enzymes in living cells. This new method was applied to induce and stop phosphatidylinositol 3-kinase activity, allowing to quantitatively measure 3,4,5-trisphosphate turnover.
AB - 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. Little helper: Chemical dimerizers are powerful tools for manipulation of enzyme activities in intact cells. The first rapidly reversible chemical dimerization system is introduced, which permits to determine kinetics of lipid metabolizing enzymes in living cells. This new method was applied to induce and stop phosphatidylinositol 3-kinase activity, allowing to quantitatively measure 3,4,5-trisphosphate turnover.
KW - FKBP
KW - PI3K
KW - SNAP
KW - reversible chemical dimerizer
UR - http://www.scopus.com/inward/record.url?scp=84903275034&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84903275034&partnerID=8YFLogxK
U2 - 10.1002/anie.201402294
DO - 10.1002/anie.201402294
M3 - Article
C2 - 24841150
AN - SCOPUS:84903275034
SN - 1433-7851
VL - 53
SP - 6720
EP - 6723
JO - Angewandte Chemie - International Edition
JF - Angewandte Chemie - International Edition
IS - 26
ER -