Skip to main content
PMC home page
Biophysical Journal logoLink to Biophysical Journal
. 2002 Jan;82(1 Pt 1):274–284. doi: 10.1016/S0006-3495(02)75393-9

Relationship of lipid rafts to transient confinement zones detected by single particle tracking.

Christian Dietrich 1, Bing Yang 1, Takahiro Fujiwara 1, Akihiro Kusumi 1, Ken Jacobson 1
PMCID: PMC1302468  PMID: 11751315

Abstract

We examined the physical and chemical characteristics of transient confinement zones (TCZs) that are detected in single particle trajectories of molecules moving within the membrane of C3H 10T1/2 murine fibroblasts and their relationship to "rafts." We studied the lateral movement of different membrane molecules thought to partition to varying degrees into or out of the putative lipid domains known as rafts. We found that lipid analogs spend significantly less time in TCZs compared with Thy-1, a glycosylphosphatidylinositol-anchored protein, and GM1, a glycosphingolipid. For Thy-1, we found that zone abundance was markedly reduced by cholesterol extraction, suggesting that a major source of the observed temporary confinement is related to the presence of raft domains. More detailed analysis of particle trajectories reveals that zones can be revisited even tens of seconds after the original escape and that diffusion within the zones is reduced by a factor of approximately 2, consistent with the zone being a cholesterol-rich liquid-ordered phase. Surprisingly, transient confinement was not strongly temperature dependent. Overall, our data demonstrate that there are raft-related domains present in certain regions of the plasma membrane of C3H cells, which can persist for tens of seconds.

Full Text

The Full Text of this article is available as a PDF (449.0 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Almeida P. F., Vaz W. L., Thompson T. E. Lateral diffusion in the liquid phases of dimyristoylphosphatidylcholine/cholesterol lipid bilayers: a free volume analysis. Biochemistry. 1992 Jul 28;31(29):6739–6747. doi: 10.1021/bi00144a013. [DOI] [PubMed] [Google Scholar]
  2. Brown D. A., London E. Functions of lipid rafts in biological membranes. Annu Rev Cell Dev Biol. 1998;14:111–136. doi: 10.1146/annurev.cellbio.14.1.111. [DOI] [PubMed] [Google Scholar]
  3. Brown D. A., London E. Structure and origin of ordered lipid domains in biological membranes. J Membr Biol. 1998 Jul 15;164(2):103–114. doi: 10.1007/s002329900397. [DOI] [PubMed] [Google Scholar]
  4. Brown D. A., Rose J. K. Sorting of GPI-anchored proteins to glycolipid-enriched membrane subdomains during transport to the apical cell surface. Cell. 1992 Feb 7;68(3):533–544. doi: 10.1016/0092-8674(92)90189-j. [DOI] [PubMed] [Google Scholar]
  5. Dietrich C., Bagatolli L. A., Volovyk Z. N., Thompson N. L., Levi M., Jacobson K., Gratton E. Lipid rafts reconstituted in model membranes. Biophys J. 2001 Mar;80(3):1417–1428. doi: 10.1016/S0006-3495(01)76114-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dráberová L., Dráber P. Thy-1 glycoprotein and src-like protein-tyrosine kinase p53/p56lyn are associated in large detergent-resistant complexes in rat basophilic leukemia cells. Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3611–3615. doi: 10.1073/pnas.90.8.3611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Heider J. G., Boyett R. L. The picomole determination of free and total cholesterol in cells in culture. J Lipid Res. 1978 May;19(4):514–518. [PubMed] [Google Scholar]
  8. Hoessli D., Rungger-Brändle E. Association of specific cell-surface glycoproteins with a triton X-100-resistant complex of plasma membrane proteins isolated from T-lymphoma cells (P1798). Exp Cell Res. 1985 Jan;156(1):239–250. doi: 10.1016/0014-4827(85)90278-2. [DOI] [PubMed] [Google Scholar]
  9. Holowka D., Sheets E. D., Baird B. Interactions between Fc(epsilon)RI and lipid raft components are regulated by the actin cytoskeleton. J Cell Sci. 2000 Mar;113(Pt 6):1009–1019. doi: 10.1242/jcs.113.6.1009. [DOI] [PubMed] [Google Scholar]
  10. Kusumi A., Sako Y. Cell surface organization by the membrane skeleton. Curr Opin Cell Biol. 1996 Aug;8(4):566–574. doi: 10.1016/s0955-0674(96)80036-6. [DOI] [PubMed] [Google Scholar]
  11. Kusumi A., Sako Y., Yamamoto M. Confined lateral diffusion of membrane receptors as studied by single particle tracking (nanovid microscopy). Effects of calcium-induced differentiation in cultured epithelial cells. Biophys J. 1993 Nov;65(5):2021–2040. doi: 10.1016/S0006-3495(93)81253-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lee G. M., Ishihara A., Jacobson K. A. Direct observation of brownian motion of lipids in a membrane. Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):6274–6278. doi: 10.1073/pnas.88.14.6274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Madore N., Smith K. L., Graham C. H., Jen A., Brady K., Hall S., Morris R. Functionally different GPI proteins are organized in different domains on the neuronal surface. EMBO J. 1999 Dec 15;18(24):6917–6926. doi: 10.1093/emboj/18.24.6917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Oliferenko S., Paiha K., Harder T., Gerke V., Schwärzler C., Schwarz H., Beug H., Günthert U., Huber L. A. Analysis of CD44-containing lipid rafts: Recruitment of annexin II and stabilization by the actin cytoskeleton. J Cell Biol. 1999 Aug 23;146(4):843–854. doi: 10.1083/jcb.146.4.843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Pralle A., Keller P., Florin E. L., Simons K., Hörber J. K. Sphingolipid-cholesterol rafts diffuse as small entities in the plasma membrane of mammalian cells. J Cell Biol. 2000 Mar 6;148(5):997–1008. doi: 10.1083/jcb.148.5.997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rodgers W., Zavzavadjian J. Glycolipid-enriched membrane domains are assembled into membrane patches by associating with the actin cytoskeleton. Exp Cell Res. 2001 Jul 15;267(2):173–183. doi: 10.1006/excr.2001.5253. [DOI] [PubMed] [Google Scholar]
  17. Saxton M. J., Jacobson K. Single-particle tracking: applications to membrane dynamics. Annu Rev Biophys Biomol Struct. 1997;26:373–399. doi: 10.1146/annurev.biophys.26.1.373. [DOI] [PubMed] [Google Scholar]
  18. Schütz G. J., Kada G., Pastushenko V. P., Schindler H. Properties of lipid microdomains in a muscle cell membrane visualized by single molecule microscopy. EMBO J. 2000 Mar 1;19(5):892–901. doi: 10.1093/emboj/19.5.892. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sheets E. D., Lee G. M., Simson R., Jacobson K. Transient confinement of a glycosylphosphatidylinositol-anchored protein in the plasma membrane. Biochemistry. 1997 Oct 14;36(41):12449–12458. doi: 10.1021/bi9710939. [DOI] [PubMed] [Google Scholar]
  20. Simons K., Ikonen E. Functional rafts in cell membranes. Nature. 1997 Jun 5;387(6633):569–572. doi: 10.1038/42408. [DOI] [PubMed] [Google Scholar]
  21. Simons K., Ikonen E. How cells handle cholesterol. Science. 2000 Dec 1;290(5497):1721–1726. doi: 10.1126/science.290.5497.1721. [DOI] [PubMed] [Google Scholar]
  22. Simson R., Sheets E. D., Jacobson K. Detection of temporary lateral confinement of membrane proteins using single-particle tracking analysis. Biophys J. 1995 Sep;69(3):989–993. doi: 10.1016/S0006-3495(95)79972-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Simson R., Yang B., Moore S. E., Doherty P., Walsh F. S., Jacobson K. A. Structural mosaicism on the submicron scale in the plasma membrane. Biophys J. 1998 Jan;74(1):297–308. doi: 10.1016/S0006-3495(98)77787-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Stahlhut M., van Deurs B. Identification of filamin as a novel ligand for caveolin-1: evidence for the organization of caveolin-1-associated membrane domains by the actin cytoskeleton. Mol Biol Cell. 2000 Jan;11(1):325–337. doi: 10.1091/mbc.11.1.325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Stauffer T. P., Meyer T. Compartmentalized IgE receptor-mediated signal transduction in living cells. J Cell Biol. 1997 Dec 15;139(6):1447–1454. doi: 10.1083/jcb.139.6.1447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tomishige M., Sako Y., Kusumi A. Regulation mechanism of the lateral diffusion of band 3 in erythrocyte membranes by the membrane skeleton. J Cell Biol. 1998 Aug 24;142(4):989–1000. doi: 10.1083/jcb.142.4.989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Varma R., Mayor S. GPI-anchored proteins are organized in submicron domains at the cell surface. Nature. 1998 Aug 20;394(6695):798–801. doi: 10.1038/29563. [DOI] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES