Interaction of glycosphingolipids and glycoproteins: thermotropic properties of model membranes containing GM1 ganglioside and glycophorin.
High-sensitivity differential scanning calorimetry (DSC) was used to study the mutual interactions between a glycoprotein (human glycophorin, GPA) and a glycosphingolipid (GM1 ganglioside) embedded in large unilamellar vesicles composed of dimyristoylphosphatidylcholine (DMPC). The DSC thermograms exhibited by DMPC/GM1 vesicles, either in the presence or in the absence of GPA, are resolvable into two components. The relative contribution of the minor component, centered at higher temperature, to the total enthalpy and its transition temperature increase with the concentration of the glycolipid embedded in the vesicles. This minor peak, undetectable in the absence of ganglioside, is indicative of the occurrence of lateral phase separation and suggests that GM1 ganglioside-enriched domains are present within the bilayer. At a given concentration of GM1 embedded in the vesicles, the proportion of the phase-separated peak is higher in the presence of GPA, suggesting that the glycoprotein enhances the tendency of GM1 to segregate. Experiments investigating the thermotropic behavior of GPA show that the temperature of irreversible thermal unfolding of the glycoprotein inserted in DMPC vesicles, centered at 65.9 degrees C in the absence of GM1, is shifted to 57.6 degrees C when GM1 is present in the bilayer. These results indicate that, at least in this experimental system, on the one hand, GPA enhances the tendency of the glycolipid to segregate within the membrane, and on the other hand, the glycolipid clusters affect the protein conformation and oligomerization in the membrane.
Department of Medical Chemistry and Biochemistry, Medical School, University of Milan, Italy.
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A
Terzaghi
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G
Tettamanti
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M
Masserini
D002152:Calorimetry, Differential Scanning; D004134:Dimyristoylphosphatidylcholine; D005677:G(M1) Ganglioside / Q000737:chemistry*; D006021:Glycophorins / Q000737:chemistry*; D006801:Humans; D066298:In Vitro Techniques; D008562:Membrane Glycoproteins / Q000737:chemistry*; D008563:Membrane Lipids / Q000737:chemistry; D008567:Membranes, Artificial; D011489:Protein Denaturation
D006021:Glycophorins; D008562:Membrane Glycoproteins; D008563:Membrane Lipids; D008567:Membranes, Artificial; D005677:G(M1) Ganglioside; D004134:Dimyristoylphosphatidylcholine