Labeling of Phosphatidylinositol Lipid Products in Cells through Metabolic Engineering by Using a Clickable myo-Inositol Probe

Tanei J. Ricks; Chelsi D. Cassilly; Adam J. Carr; Daiane S. Alves; Shahrina Alam; Kathrin Tscherch; Timothy W. Yokley; Cameron E. Workman; Jennifer L. Morrell-Falvey; Francisco N. Barrera; Todd B. Reynolds; Michael D. Best

doi:10.1002/cbic.201800248

Labeling of Phosphatidylinositol Lipid Products in Cells through Metabolic Engineering by Using a Clickable myo-Inositol Probe

Tanei J. Ricks, Chelsi D. Cassilly, Adam J. Carr, Daiane S. Alves, Shahrina Alam, Kathrin Tscherch, Timothy W. Yokley, Cameron E. Workman, Jennifer L. Morrell-Falvey, Francisco N. Barrera, Todd B. Reynolds, Michael D. Best

Molecular and Cellular Imaging

Research output: Contribution to journal › Article › peer-review

27 Scopus citations

Abstract

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.

Original language	English
Pages (from-to)	172-180
Number of pages	9
Journal	ChemBioChem
Volume	20
Issue number	2
DOIs	https://doi.org/10.1002/cbic.201800248
State	Published - Jan 18 2019

Funding

This research was supported by funding from the National Institutes of Health (R15GM120705). A portion of the fluorescence imaging studies in this work was conducted at the Center for Nanophase Materials Sciences, which is a DOE Office of Science User Facility.

Keywords

click chemistry
fluorescent probes
lipids
metabolic labeling
phospholipids

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10.1002/cbic.201800248

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Ricks, T. J., Cassilly, C. D., Carr, A. J., Alves, D. S., Alam, S., Tscherch, K., Yokley, T. W., Workman, C. E., Morrell-Falvey, J. L., Barrera, F. N., Reynolds, T. B., & Best, M. D. (2019). Labeling of Phosphatidylinositol Lipid Products in Cells through Metabolic Engineering by Using a Clickable myo-Inositol Probe. ChemBioChem, 20(2), 172-180. https://doi.org/10.1002/cbic.201800248

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abstract = "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.",

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AU - Reynolds, Todd B.

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Labeling of Phosphatidylinositol Lipid Products in Cells through Metabolic Engineering by Using a Clickable myo-Inositol Probe

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