Lo/Ld phase coexistence modulation induced by GM1

• Published in: Biochimica et biophysica acta 2014-08, Vol.1838 (8), p.2105-2114
• Main Authors: Puff, Nicolas, Watanabe, Chiho, Seigneuret, Michel, Angelova, Miglena I., Staneva, Galya
• Format: Article
• Language: English
• Subjects: 2-Naphthylamine - analogs & derivatives; Biological Physics; Cell Membrane - chemistry; Fluorescent Dyes; G(M1) Ganglioside - chemistry; GM1; GUV; Laurates; Laurdan; Lipid Bilayers - chemistry; Lo/Ld phase coexistence; Membrane Microdomains - chemistry; Microdomain; Nanoscale domain; Physics; Spectrometry, Fluorescence; Unilamellar Liposomes - chemistry
• ISSN: 0005-2736; 0006-3002; 1879-2642
• DOI: 10.1016/j.bbamem.2014.05.002

Lipid rafts are assumed to undergo biologically important size-modulations from nanorafts to microrafts. Due to the complexity of cellular membranes, model systems become important tools, especially for the investigation of the factors affecting “raft-like” Lo domain size and the search for Lo nanodomains as precursors in Lo microdomain formation. Because lipid compositional change is the primary mechanism by which a cell can alter membrane phase behavior, we studied the effect of the ganglioside GM1 concentration on the Lo/Ld lateral phase separation in PC/SM/Chol/GM1 bilayers. GM1 above 1mol % abolishes the formation of the micrometer-scale Lo domains observed in GUVs. However, the apparently homogeneous phase observed in optical microscopy corresponds in fact, within a certain temperature range, to a Lo/Ld lateral phase separation taking place below the optical resolution. This nanoscale phase separation is revealed by fluorescence spectroscopy, including C12NBD-PC self-quenching and Laurdan GP measurements, and is supported by Gaussian spectral decomposition analysis. The temperature of formation of nanoscale Lo phase domains over an Ld phase is determined, and is shifted to higher values when the GM1 content increases. A “morphological” phase diagram could be made, and it displays three regions corresponding respectively to Lo/Ld micrometric phase separation, Lo/Ld nanometric phase separation, and a homogeneous Ld phase. We therefore show that a lipid only-based mechanism is able to control the existence and the sizes of phase-separated membrane domains. GM1 could act on the line tension, “arresting” domain growth and thereby stabilizing Lo nanodomains. [Display omitted] •The effect of GM1 concentration on the Lo/Ld lateral phase separation was studied.•GM1 above 1mol % abolishes the formation of the micrometer-scale Lo domains.•For a certain temperature range, a Lo/Ld nanoscale phase separation is revealed.•The demixing temperature is shifted to a higher value when the GM1 content increases.•A lipid only-based mechanism is able to modulate phase-separated membrane domains.