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Originally published as MBC in Press, 10.1091/mbc.E07-04-0330 on October 17, 2007

Vol. 18, Issue 12, 5113-5123, December 2007

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Endocytic Trafficking of Sphingomyelin Depends on Its Acyl Chain LengthFormula

Mirkka Koivusalo*,{dagger}, Maurice Jansen*, Pentti Somerharju{ddagger}, and Elina Ikonen*

*Institute of Biomedicine/Anatomy and {ddagger}Institute of Biomedicine/Biochemistry, University of Helsinki, FIN-00014, Helsinki, Finland

Submitted April 12, 2007; Revised September 21, 2007; Accepted October 5, 2007
Monitoring Editor: Howard Riezman

To study the principles of endocytic lipid trafficking, we introduced pyrene sphingomyelins (PyrSMs) with varying acyl chain lengths and domain partitioning properties into human fibroblasts or HeLa cells. We found that a long-chain, ordered-domain preferring PyrSM was targeted Hrs and Tsg101 dependently to late endosomal compartments and recycled to the plasma membrane in an NPC1- and cholesterol-dependent manner. A short-chain, disordered domain preferring PyrSM recycled more effectively, by using Hrs-, Tsg101- and NPC1-independent routing that was insensitive to cholesterol loading. Similar chain length-dependent recycling was observed for unlabeled sphingomyelins (SMs). The findings 1) establish acyl chain length as an important determinant in the endocytic trafficking of SMs, 2) implicate ESCRT complex proteins and NPC1 in the endocytic recycling of ordered domain lipids to the plasma membrane, and 3) introduce long-chain PyrSM as the first fluorescent lipid tracing this pathway.


This article was published online ahead of print in MBC in Press (http://www.molbiolcell.org/cgi/doi/10.1091/mbc.E07-04-0330) on October 17, 2007.

Formula The online version of this article contains supplemental material at MBC Online (http://www.molbiolcell.org).

{dagger} Present address: Programme in Cell Biology, Hospital for Sick Children, 555 University Ave., Toronto, ON, M5G 1X8, Canada.

Address correspondence to: Elina Ikonen (elina.ikonen{at}helsinki.fi).

Abbreviations used: BMP, bis(monoacylglycero)phosphate; CD, cyclodextrin; DRM, detergent-resistant membrane; ESCRT, endosomal sorting complex required for transport; ESI-MS, electrospray ionization mass spectrometry; Hrs, hepatocyte growth factor receptor substrate; MVB, multivesicular body; NPA, Niemann-Pick type A; NPC, Niemann-Pick type C; PM, plasma membrane; PyrSM, pyrenylacylsphingomyelin; SM, sphingomyelin; SMase, sphingomyelinase; TNP-LPE, trinitrophenyl-lysophosphatidylethanolamine; Tsg, tumor susceptibility gene.







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