Atomic Force Microscopy Studies of Ganglioside GM1 Domains in Phosphatidylcholine and Phosphatidylcholine/Cholesterol Bilayers

The distribution of ganglioside in supported lipid bilayers has been studied by atomic force microscopy. Hybrid dipalmitoylphosphatidylcholine (DPPC)/dipalmitoylphosphatidylethanolamine (DPPE) and (2:1 DPPC/cholesterol)/DPPE bilayers were prepared using the Langmuir Blodgett technique. Egg PC and DP...

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Bibliographic Details
Published in:Biophysical journal 2001-08, Vol.81 (2), p.1059-1069
Main Authors: Yuan, Chunbo, Johnston, Linda J.
Format: Article
Language:English
Subjects:
1,2-Dipalmitoylphosphatidylcholine - metabolism
Cholera Toxin - metabolism
Cholesterol - metabolism
G(M1) Ganglioside - metabolism
Lipid Bilayers - chemistry
Lipid Bilayers - metabolism
Membrane Fusion
Membrane Microdomains
Microscopy, Atomic Force
Phosphatidylcholines - metabolism
Phosphatidylethanolamines - metabolism
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Summary:The distribution of ganglioside in supported lipid bilayers has been studied by atomic force microscopy. Hybrid dipalmitoylphosphatidylcholine (DPPC)/dipalmitoylphosphatidylethanolamine (DPPE) and (2:1 DPPC/cholesterol)/DPPE bilayers were prepared using the Langmuir Blodgett technique. Egg PC and DPPC bilayers were prepared by vesicle fusion. Addition of ganglioside GM1 to each of the lipid bilayers resulted in the formation of heterogeneous surfaces that had numerous small raised domains (30–200 nm in diameter). Incubation of these bilayers with cholera toxin B subunit resulted in the detection of small protein aggregates, indicating specific binding of the protein to the GM1-rich microdomains. Similar results were obtained for DPPC, DPPC/cholesterol, and egg PC, demonstrating that the overall bilayer morphology was not dependent on the method of bilayer preparation or the fluidity of the lipid mixture. However, bilayers produced by vesicle fusion provided evidence for asymmetrically distributed GM1 domains that probably reflect the presence of ganglioside in both inner and outer monolayers of the initial vesicle. The results are discussed in relation to recent inconsistencies in the estimation of sizes of lipid rafts in model and natural membranes. It is hypothesized that small ganglioside-rich microdomains may exist within larger ordered domains in both natural and model membranes.
ISSN:0006-3495
1542-0086
DOI:10.1016/S0006-3495(01)75763-3