Probing lipid mobility of raft-exhibiting model membranes by fluorescence correlation spectroscopy

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • Nicoletta Kahya - (Author)
  • Dag Scherfeld - (Author)
  • Kirsten Bacia - (Author)
  • Bert Poolman - (Author)
  • Petra Schwille - , Chair of Biophysics (Author)

Abstract

Confocal fluorescence microscopy and fluorescence correlation spectroscopy (FCS) have been employed to investigate the lipid spatial and dynamic organization in giant unilamellar vesicles (GUVs) prepared from ternary mixtures of dioleoyl-phosphatidylcholine/sphingo-myelin/cholesterol. For a certain range of cholesterol concentration, formation of domains with raft-like properties was observed. Strikingly, the lipophilic probe 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI-C18) was excluded from sphingomyelin-enriched regions, where the raft marker ganglioside GM1 was localized. Cholesterol was shown to promote lipid segregation in dioleoyl-phosphatidylcholine-enriched, liquid -disordered, and sphingomyelin-enriched, liquid-ordered phases. Most importantly, the lipid mobility in sphingomyelin-enriched regions significantly increased by increasing the cholesterol concentration. These results pinpoint the key role, played by cholesterol in tuning lipid dynamics in membranes. At cholesterol concentrations > 50 mol%, domains vanished and the lipid diffusion slowed down upon further addition of cholesterol. By taking the molecular diffusion coefficients as a fingerprint of membrane phase compositions, FCS is proven to evaluate domain lipid compositions. Moreover, FCS data from ternary and binary mixtures have been used to build a ternary phase diagram, which shows areas of phase coexistence, transition points, and, importantly, how lipid dynamics varies f between and within phase regions.

Details

Original languageEnglish
Pages (from-to)28109-28115
Number of pages7
JournalJournal of Biological Chemistry
Volume278
Issue number30
Publication statusPublished - 25 Jul 2003
Peer-reviewedYes

External IDs

PubMed 12736276

Keywords