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Fluorescence Resonance Energy Transfer between Lipid Probes Detects Nanoscopic Heterogeneity in the Plasma Membrane of Live Cells
- Source :
- Biophysical Journal. 92(10):3564-3574
- Publication Year :
- 2007
- Publisher :
- Elsevier BV, 2007.
-
Abstract
- Fluorescence resonance energy transfer (FRET) between matched carbocyanine lipid analogs in the plasma membrane outer leaflet of RBL mast cells was used to investigate lateral distributions of lipids and to develop a general method for quantitative measurements of lipid heterogeneity in live cell membranes. FRET measured as fluorescence quenching of long-chain donor probes such as DiO-C18 is greater with long-chain, saturated acceptor probes such as DiI-C16 than with unsaturated or shorter-chain acceptors with the same chromophoric headgroup compared at identical concentrations. FRET measurements between these lipid probes in model membranes support the conclusion that differential donor quenching is not caused by nonideal mixing or spectroscopic differences. Sucrose gradient analysis of plasma membrane-labeled, Triton X-100-lysed cells shows that proximity measured by FRET correlates with the extent of lipid probe partitioning into detergent-resistant membranes. FRET between DiO-C16 and DiI-C16 is sensitive to cholesterol depletion and disruption of liquid order (Lo) by short-chain ceramides, and it is enhanced by cross linking of Lo-associated proteins. Consistent results are obtained when homo-FRET is measured by decreased fluorescence anisotropy of DiI-C16. These results support the existence of nanometer-scale Lo/liquid disorder heterogeneity of lipids in the outer leaflet of the plasma membrane in live cells.
- Subjects :
- Membrane Fluidity
Membrane lipids
Analytical chemistry
Biophysics
7. Clean energy
Cell Line
03 medical and health sciences
Membrane Lipids
Membrane fluidity
Fluorescence Resonance Energy Transfer
Animals
Mast Cells
030304 developmental biology
Fluorescent Dyes
0303 health sciences
Liposome
Quenching (fluorescence)
Membranes
Chemistry
030302 biochemistry & molecular biology
Acceptor
Nanostructures
Rats
Förster resonance energy transfer
Membrane
Liposomes
lipids (amino acids, peptides, and proteins)
Cell Surface Extensions
Fluorescence anisotropy
Subjects
Details
- ISSN :
- 00063495
- Volume :
- 92
- Issue :
- 10
- Database :
- OpenAIRE
- Journal :
- Biophysical Journal
- Accession number :
- edsair.doi.dedup.....6ba97d4f6b9132ab9b43844565497cff
- Full Text :
- https://doi.org/10.1529/biophysj.106.094730