关键词: FKBP PI3K PIP3 SNAP reversible chemical dimerizer

Mesh : Dimerization HeLa Cells Humans Kinetics Phosphatidylinositol 3-Kinases / metabolism Phosphatidylinositol Phosphates / metabolism Phosphorylation Proto-Oncogene Proteins c-akt / metabolism Signal Transduction / drug effects Tacrolimus / pharmacology

来  源:   DOI:10.1002/anie.201402294   PDF(Sci-hub)

Abstract:
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.
摘要:
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