期刊
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
卷 53, 期 26, 页码 6720-6723出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201402294
关键词
FKBP; PI3K; PIP3; reversible chemical dimerizer; SNAP
资金
- DFG [Transregio 83]
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 (PIP3) and its downstream effectors by confocal fluorescence microscopy as well as standard biochemical methods.
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