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Title: 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:
 [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1]; ORCiD logo [2];  [1];  [1]; ORCiD logo [1]
  1. Univ. of Tennessee, Knoxville, TN (United States)
  2. 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. 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., and Best, Michael D. Sat . "Labeling of Phosphatidylinositol Lipid Products in Cells through Metabolic Engineering by Using a Clickable myo-Inositol Probe". United States. doi:10.1002/cbic.201800248.
@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 = {2018},
month = {8}
}

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Works referenced in this record:

Bioorthogonal Chemistry: Fishing for Selectivity in a Sea of Functionality
journal, September 2009

  • Sletten, Ellen?M.; Bertozzi, Carolyn?R.
  • Angewandte Chemie International Edition, Vol. 48, Issue 38, p. 6974-6998
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