Direct chemical evidence for sphingolipid domains in the plasma membranes of fibroblasts [High-Resolution Chemical Imaging of Sphingolipid Distribution in the Plasma Membrane]

Frisz, Jessica F.; Lou, Kaiyan; Klitzing, Haley A.; Hanafin, William P.; Lizunov, Vladimir; Wilson, Robert L.; Carpenter, Kevin J.; Kim, Raehyun; Hutcheon, Ian D.; Zimmerberg, Joshua; Weber, Peter K.; Kraft, Mary L.

doi:10.1073/pnas.1216585110

Title: Direct chemical evidence for sphingolipid domains in the plasma membranes of fibroblasts [High-Resolution Chemical Imaging of Sphingolipid Distribution in the Plasma Membrane]

Journal Article · Mon Jan 28 00:00:00 EST 2013 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.1216585110· OSTI ID:1396208

Frisz, Jessica F. ^[1]; Lou, Kaiyan ^[1]; Klitzing, Haley A. ^[1]; Hanafin, William P. ^[1]; Lizunov, Vladimir ^[2]; Wilson, Robert L. ^[1]; Carpenter, Kevin J. ^[3]; Kim, Raehyun ^[1]; Hutcheon, Ian D. ^[3]; Zimmerberg, Joshua ^[2]; Weber, Peter K. ^[3]; Kraft, Mary L. ^[1]

Univ. of Illinois, Urbana, IL (United States). School of Chemical Sciences
National Inst. of Health, Bethesda, MD (United States). Program in Physical Biology, Eunice Kennedy Shriver National Inst. of Child Health and Human Development
Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States). Glenn T. Seaborg Inst.

Sphingolipids play important roles in plasma membrane structure and cell signaling. Yet, their lateral distribution in the plasma membrane is poorly understood. Here we quantitatively analyzed the sphingolipid organization on the entire dorsal surface of intact cells by mapping the distribution of ¹⁵N-enriched ions from metabolically labeled ¹⁵N-sphingolipids in the plasma membrane using high-resolution imaging mass spectrometry. Many types of control experiments (internal, positive, negative, and fixation temperature), along with parallel experiments involving the imaging of fluorescent sphingolipids$$-$$both in living cells and during fixation of living cells$$-$$exclude potential artifacts. Micrometer-scale sphingolipid patches consisting of numerous ¹⁵Nsphingolipid microdomains with mean diameters of ~200 nm are always present in the plasma membrane. Depletion of 30% of the cellular cholesterol did not eliminate the sphingolipid domains, but did reduce their abundance and long range organization in the plasma membrane. In contrast, disruption of the cytoskeleton eliminated the sphingolipid domains. These results indicate that these sphingolipid assemblages are not lipid rafts, and are instead a distinctly different type of sphingolipid-enriched plasma membrane domain that depends upon cortical actin.

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Research Organization:: Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States)

Sponsoring Organization:: USDOE; National Institutes of Health (NIH); National Science Foundation (NSF)

Grant/Contract Number:: AC52-07NA27344; FG02-07ER4647; CHE-1058809

OSTI ID:: 1396208

Report Number(s):: LLNL-JRNL-463256

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Vol. 110, Issue 8; ISSN 0027-8424

Publisher:: National Academy of Sciences, Washington, DC (United States)Copyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 145 works

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Title: Direct chemical evidence for sphingolipid domains in the plasma membranes of fibroblasts [High-Resolution Chemical Imaging of Sphingolipid Distribution in the Plasma Membrane]

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