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Labeling of Phosphatidylinositol Lipid Products in Cells through Metabolic Engineering by Using a Clickable myo ‐Inositol Probe
Author(s) -
Ricks Tanei J.,
Cassilly Chelsi D.,
Carr Adam J.,
Alves Daiane S.,
Alam Shahrina,
Tscherch Kathrin,
Yokley Timothy W.,
Workman Cameron E.,
MorrellFalvey Jennifer L.,
Barrera Francisco N.,
Reynolds Todd B.,
Best Michael D.
Publication year - 2019
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201800248
Subject(s) - bioorthogonal chemistry , inositol , phosphatidylinositol , biochemistry , biosynthesis , chemistry , metabolic engineering , fluorescence microscope , click chemistry , microbiology and biotechnology , biophysics , fluorescence , biology , signal transduction , enzyme , receptor , combinatorial chemistry , physics , quantum mechanics
Phosphatidylinositol (PI) lipids control critical biological processes, so aberrant biosynthesis often leads to disease. As a result, the capability to track the production and localization of these compounds in cells is vital for elucidating their complex roles. Herein, we report the design, synthesis, and application of clickable myo ‐inositol probe 1 a for bioorthogonal labeling of PI products. To validate this platform, we initially conducted PI synthase assays to show that 1 a inhibits PI production in vitro. Fluorescence microscopy experiments next showed probe‐dependent imaging in T‐24 human bladder cancer and Candida albicans cells. Growth studies in the latter showed that replacement of myo ‐inositol with probe 1 a led to an enhancement in cell growth. Finally, fluorescence‐based TLC analysis and mass spectrometry experiments support the labeling of PI lipids. This approach provides a promising means for tracking the complex biosynthesis and trafficking of these lipids in cells.