Labeling of Phosphatidylinositol Lipid Products in Cells through Metabolic Engineering by Using a Clickable myo-Inositol Probe
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. In this paper, 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. Lastly, 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.
- Authors:
-
[2];
[1]
- Univ. of Tennessee, Knoxville, TN (United States)
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
- Publication Date:
- Research Org.:
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1494897
- Grant/Contract Number:
- AC05-00OR22725
- Resource Type:
- Accepted Manuscript
- Journal Name:
- ChemBioChem: a European journal of chemical biology
- Additional Journal Information:
- Journal Volume: 20; Journal Issue: 2; Journal ID: ISSN 1439-4227
- Publisher:
- ChemPubSoc Europe
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; click chemistry; fluorescent probes; lipids; metabolic labeling; phospholipids
Citation Formats
Ricks, Tanei, Cassilly, Chelsi D., Carr, Adam J., Alves, Daiane S., Alam, Shahrina, Tscherch, Kathrin, Yokley, Timothy W., Workman, Cameron E., Morrell-Falvey, Jennifer L., Barrera, Francisco N., Reynolds, Todd B., and Best, Michael D. Labeling of Phosphatidylinositol Lipid Products in Cells through Metabolic Engineering by Using a Clickable myo-Inositol Probe. United States: N. p., 2018.
Web. doi:10.1002/cbic.201800248.
Ricks, Tanei, Cassilly, Chelsi D., Carr, Adam J., Alves, Daiane S., Alam, Shahrina, Tscherch, Kathrin, Yokley, Timothy W., Workman, Cameron E., Morrell-Falvey, Jennifer L., Barrera, Francisco N., Reynolds, Todd B., & Best, Michael D. Labeling of Phosphatidylinositol Lipid Products in Cells through Metabolic Engineering by Using a Clickable myo-Inositol Probe. United States. https://doi.org/10.1002/cbic.201800248
Ricks, Tanei, Cassilly, Chelsi D., Carr, Adam J., Alves, Daiane S., Alam, Shahrina, Tscherch, Kathrin, Yokley, Timothy W., Workman, Cameron E., Morrell-Falvey, Jennifer L., Barrera, Francisco N., Reynolds, Todd B., and Best, Michael D. Sat .
"Labeling of Phosphatidylinositol Lipid Products in Cells through Metabolic Engineering by Using a Clickable myo-Inositol Probe". United States. https://doi.org/10.1002/cbic.201800248. https://www.osti.gov/servlets/purl/1494897.
@article{osti_1494897,
title = {Labeling of Phosphatidylinositol Lipid Products in Cells through Metabolic Engineering by Using a Clickable myo-Inositol Probe},
author = {Ricks, Tanei and Cassilly, Chelsi D. and Carr, Adam J. and Alves, Daiane S. and Alam, Shahrina and Tscherch, Kathrin and Yokley, Timothy W. and Workman, Cameron E. and Morrell-Falvey, Jennifer L. and Barrera, Francisco N. and Reynolds, Todd B. and Best, Michael D.},
abstractNote = {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. In this paper, 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. Lastly, 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.},
doi = {10.1002/cbic.201800248},
journal = {ChemBioChem: a European journal of chemical biology},
number = 2,
volume = 20,
place = {United States},
year = {Sat Aug 11 00:00:00 EDT 2018},
month = {Sat Aug 11 00:00:00 EDT 2018}
}
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