skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Sphingolipid domains in the plasma membranes of fibroblasts are not enriched with cholesterol

Abstract

The plasma membranes of mammalian cells are widely expected to contain domains that are enriched with cholesterol and sphingolipids. In this work, we have used high-resolution secondary ion mass spectrometry to directly map the distributions of isotope-labeled cholesterol and sphingolipids in the plasma membranes of intact fibroblast cells. Although acute cholesterol depletion reduced sphingolipid domain abundance, cholesterol was evenly distributed throughout the plasma membrane and was not enriched within the sphingolipid domains. As a result, we rule out favorable cholesterol-sphingolipid interactions as dictating plasma membrane organization in fibroblast cells. Because the sphingolipid domains are disrupted by drugs that depolymerize the cells actin cytoskeleton, cholesterol must instead affect the sphingolipid organization via an indirect mechanism that involves the cytoskeleton.

Authors:
 [1];  [1];  [1];  [2];  [2];  [3];  [4]
  1. Univ. of Illinois, Urbana, IL (United States)
  2. Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
  3. National Institutes of Health, Bethesda, MD (United States)
  4. Univ. of Illinois, Urbana, IL (United States); Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
Publication Date:
Research Org.:
Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1313553
Report Number(s):
LLNL-JRNL-638518
Journal ID: ISSN 0021-9258
Grant/Contract Number:  
AC52-07NA27344
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Journal of Biological Chemistry
Additional Journal Information:
Journal Volume: 288; Journal Issue: 23; Journal ID: ISSN 0021-9258
Publisher:
American Society for Biochemistry and Molecular Biology
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL AND ANALYTICAL CHEMISTRY; cholesterol; isotopic tracers; lipid raft; mass spectrometry (MS); membrane structure; plasma membrane; sphingolipid

Citation Formats

Frisz, Jessica F., Klitzing, Haley A., Lou, Kaiyan, Hutcheon, Ian D., Weber, Peter K., Zimmerberg, Joshua, and Kraft, Mary L. Sphingolipid domains in the plasma membranes of fibroblasts are not enriched with cholesterol. United States: N. p., 2013. Web. doi:10.1074/jbc.M113.473207.
Frisz, Jessica F., Klitzing, Haley A., Lou, Kaiyan, Hutcheon, Ian D., Weber, Peter K., Zimmerberg, Joshua, & Kraft, Mary L. Sphingolipid domains in the plasma membranes of fibroblasts are not enriched with cholesterol. United States. doi:10.1074/jbc.M113.473207.
Frisz, Jessica F., Klitzing, Haley A., Lou, Kaiyan, Hutcheon, Ian D., Weber, Peter K., Zimmerberg, Joshua, and Kraft, Mary L. Mon . "Sphingolipid domains in the plasma membranes of fibroblasts are not enriched with cholesterol". United States. doi:10.1074/jbc.M113.473207. https://www.osti.gov/servlets/purl/1313553.
@article{osti_1313553,
title = {Sphingolipid domains in the plasma membranes of fibroblasts are not enriched with cholesterol},
author = {Frisz, Jessica F. and Klitzing, Haley A. and Lou, Kaiyan and Hutcheon, Ian D. and Weber, Peter K. and Zimmerberg, Joshua and Kraft, Mary L.},
abstractNote = {The plasma membranes of mammalian cells are widely expected to contain domains that are enriched with cholesterol and sphingolipids. In this work, we have used high-resolution secondary ion mass spectrometry to directly map the distributions of isotope-labeled cholesterol and sphingolipids in the plasma membranes of intact fibroblast cells. Although acute cholesterol depletion reduced sphingolipid domain abundance, cholesterol was evenly distributed throughout the plasma membrane and was not enriched within the sphingolipid domains. As a result, we rule out favorable cholesterol-sphingolipid interactions as dictating plasma membrane organization in fibroblast cells. Because the sphingolipid domains are disrupted by drugs that depolymerize the cells actin cytoskeleton, cholesterol must instead affect the sphingolipid organization via an indirect mechanism that involves the cytoskeleton.},
doi = {10.1074/jbc.M113.473207},
journal = {Journal of Biological Chemistry},
issn = {0021-9258},
number = 23,
volume = 288,
place = {United States},
year = {2013},
month = {4}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Citation Metrics:
Cited by: 58 works
Citation information provided by
Web of Science

Save / Share:

Works referenced in this record:

Intracellular sterol transport and distribution
journal, August 2006


Do Lipid Rafts Exist?
journal, November 2011


Lipid Rafts As a Membrane-Organizing Principle
journal, December 2009


Revitalizing membrane rafts: new tools and insights
journal, September 2010

  • Simons, Kai; Gerl, Mathias J.
  • Nature Reviews Molecular Cell Biology, Vol. 11, Issue 10
  • DOI: 10.1038/nrm2977

Direct observation of the nanoscale dynamics of membrane lipids in a living cell
journal, December 2008

  • Eggeling, Christian; Ringemann, Christian; Medda, Rebecca
  • Nature, Vol. 457, Issue 7233
  • DOI: 10.1038/nature07596

Quantitative imaging of cells with multi-isotope imaging mass spectrometry (MIMS)—Nanoautography with stable isotope tracers
journal, July 2006

  • McMahon, Greg; Glassner, Brian J.; Lechene, Claude P.
  • Applied Surface Science, Vol. 252, Issue 19, p. 6895-6906
  • DOI: 10.1016/j.apsusc.2006.02.170

Correlated AFM and NanoSIMS imaging to probe cholesterol-induced changes in phase behavior and non-ideal mixing in ternary lipid membranes
journal, January 2011

  • Anderton, Christopher R.; Lou, Kaiyan; Weber, Peter K.
  • Biochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 1808, Issue 1
  • DOI: 10.1016/j.bbamem.2010.09.016

Multi-isotope imaging mass spectrometry quantifies stem cell division and metabolism
journal, January 2012

  • Steinhauser, Matthew L.; Bailey, Andrew P.; Senyo, Samuel E.
  • Nature, Vol. 481, Issue 7382
  • DOI: 10.1038/nature10734

Direct chemical evidence for sphingolipid domains in the plasma membranes of fibroblasts
journal, January 2013

  • Frisz, J. F.; Lou, K.; Klitzing, H. A.
  • Proceedings of the National Academy of Sciences, Vol. 110, Issue 8
  • DOI: 10.1073/pnas.1216585110

Fluorinated Colloidal Gold Immunolabels for Imaging Select Proteins in Parallel with Lipids Using High-Resolution Secondary Ion Mass Spectrometry
journal, February 2012

  • Wilson, Robert L.; Frisz, Jessica F.; Hanafin, William P.
  • Bioconjugate Chemistry, Vol. 23, Issue 3
  • DOI: 10.1021/bc200482z

Quantitative electron microscopy and fluorescence spectroscopy of the membrane distribution of influenza hemagglutinin
journal, June 2005

  • Hess, Samuel T.; Kumar, Mukesh; Verma, Anil
  • The Journal of Cell Biology, Vol. 169, Issue 6
  • DOI: 10.1083/jcb.200412058

Progressive ordering with decreasing temperature of the phospholipids of influenza virus
journal, March 2008

  • Polozov, Ivan V.; Bezrukov, Ludmila; Gawrisch, Klaus
  • Nature Chemical Biology, Vol. 4, Issue 4
  • DOI: 10.1038/nchembio.77

Functional rafts in cell membranes
journal, June 1997

  • Simons, Kai; Ikonen, Elina
  • Nature, Vol. 387, Issue 6633
  • DOI: 10.1038/42408

A stereodivergent synthesis of D-erythro-sphingosine and D-threo-sphingosine from L-serine
journal, September 1988

  • Garner, Philip; Park, Jung Min; Malecki, Elise
  • The Journal of Organic Chemistry, Vol. 53, Issue 18
  • DOI: 10.1021/jo00253a039

Gold-Enhanced Biomolecular Surface Imaging of Cells and Tissue by SIMS and MALDI Mass Spectrometry
journal, February 2006

  • Altelaar, A. F. Maarten; Klinkert, Ivo; Jalink, Kees
  • Analytical Chemistry, Vol. 78, Issue 3
  • DOI: 10.1021/ac0513111

Imaging and 3D Elemental Characterization of Intact Bacterial Spores by High-Resolution Secondary Ion Mass Spectrometry
journal, August 2008

  • Ghosal, Sutapa; Fallon, Stewart J.; Leighton, Terrance J.
  • Analytical Chemistry, Vol. 80, Issue 15
  • DOI: 10.1021/ac8006279

Quantitative Imaging of Nitrogen Fixation by Individual Bacteria Within Animal Cells
journal, September 2007


An adhesion-based method for plasma membrane isolation: Evaluating cholesterol extraction from cells and their membranes
journal, November 2009

  • Bezrukov, Ludmila; Blank, Paul S.; Polozov, Ivan V.
  • Analytical Biochemistry, Vol. 394, Issue 2
  • DOI: 10.1016/j.ab.2009.07.027

Miscibility Phase Diagrams of Giant Vesicles Containing Sphingomyelin
journal, April 2005


Colocalization of the Ganglioside G M1 and Cholesterol Detected by Secondary Ion Mass Spectrometry
journal, April 2013

  • Lozano, Mónica M.; Liu, Zhao; Sunnick, Eva
  • Journal of the American Chemical Society, Vol. 135, Issue 15
  • DOI: 10.1021/ja310831m

Use of cyclodextrins to manipulate plasma membrane cholesterol content: Evidence, misconceptions and control strategies
journal, June 2007

  • Zidovetzki, Raphael; Levitan, Irena
  • Biochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 1768, Issue 6
  • DOI: 10.1016/j.bbamem.2007.03.026

Modification of Cellular Cholesterol Content Affects Traction Force, Adhesion and Cell Spreading
journal, March 2010

  • Norman, Leann L.; Oetama, Ratna J.; Dembo, Micah
  • Cellular and Molecular Bioengineering, Vol. 3, Issue 2
  • DOI: 10.1007/s12195-010-0119-x

Membrane cholesterol, lateral mobility, and the phosphatidylinositol 4,5-bisphosphate-dependent organization of cell actin
journal, November 2003

  • Kwik, J.; Boyle, S.; Fooksman, D.
  • Proceedings of the National Academy of Sciences, Vol. 100, Issue 24
  • DOI: 10.1073/pnas.2336102100

Changes in cholesterol levels in the plasma membrane modulate cell signaling and regulate cell adhesion and migration on fibronectin
journal, March 2007

  • Ramprasad, O. G.; Srinivas, G.; Rao, K. Sridhar
  • Cell Motility and the Cytoskeleton, Vol. 64, Issue 3
  • DOI: 10.1002/cm.20176

Cholesterol modulates cell signaling and protein networking by specifically interacting with PDZ domain-containing scaffold proteins
journal, January 2012

  • Sheng, Ren; Chen, Yong; Yung Gee, Heon
  • Nature Communications, Vol. 3, Issue 1
  • DOI: 10.1038/ncomms2221

OSBP Is a Cholesterol-Regulated Scaffolding Protein in Control of ERK 1/2 Activation
journal, March 2005


Nanoclusters of GPI-Anchored Proteins Are Formed by Cortical Actin-Driven Activity
journal, December 2008


Active Remodeling of Cortical Actin Regulates Spatiotemporal Organization of Cell Surface Molecules
journal, June 2012


Domain Formation in the Plasma Membrane: Roles of Nonequilibrium Lipid Transport and Membrane Proteins
journal, April 2008


Nonequilibrium Raftlike Membrane Domains under Continuous Recycling
journal, October 2005


Dynamic Patches of Membrane Proteins
journal, September 2007


A Model for Membrane Patchiness: Lateral Diffusion in the Presence of Barriers and Vesicle Traffic
journal, December 1999


    Works referencing / citing this record:

    Regulation of monoamine transporters and receptors by lipid microdomains: implications for depression
    journal, June 2018

    • Liu, Joanne J.; Hezghia, Adrienne; Shaikh, Saame Raza
    • Neuropsychopharmacology, Vol. 43, Issue 11
    • DOI: 10.1038/s41386-018-0133-6

    Regulation of monoamine transporters and receptors by lipid microdomains: implications for depression
    journal, June 2018

    • Liu, Joanne J.; Hezghia, Adrienne; Shaikh, Saame Raza
    • Neuropsychopharmacology, Vol. 43, Issue 11
    • DOI: 10.1038/s41386-018-0133-6

    Mesoscale organization of domains in the plasma membrane – beyond the lipid raft
    journal, February 2018


    Interleaflet Coupling, Pinning, and Leaflet Asymmetry—Major Players in Plasma Membrane Nanodomain Formation
    journal, January 2017