The role of protein-linked oligosaccharide in the bilayer stabilization activity of glycophorin A for dioleoylphosphatidylethanolamine liposomes.
The importance of the protein-linked carbohydrates for the stabilization of dioleoylphosphatidylethanolamine (DOPE) bilayers has been investigated using glycophorin A, the major sialoglycoprotein of the human erythrocyte membrane, as a stabilizer. Two major types of glycophorin, differing in the sialic acid content, were used in the study. Type MM contains 19.2 +/- 2.5 sialic residues per molecule of glycophorin, and type NN contains 10.8 +/- 1.2. Type MM could stabilize DOPE bilayers at 0.5 mol%, whereas type NN was unable to do so even at 1 mol%. The importance of the sialic acid content to the stabilization activity of glycophorin was further confirmed by the observation that the neuraminidase-treated type MM showed a lower stabilization activity than the untreated type. Since type NN had no stabilizing activity, we attempted to couple a trisaccharide, NeuNAc----Gal----Glc, to type NN by reductive amination. 2.5 +/- 0.8 saccharide chains were added per molecule of type NN. The trisaccharide-attached type NN showed a greater stabilization activity than the parent type NN molecule, indicating again that the sialic acid content of the stabilizer molecule determines the stabilization activity. Addition of wheat-germ agglutinin (WGA), which binds to the sialic acid residues of a glycoprotein, to type MM-stabilized liposomes caused rapid aggregation and destabilization of liposomes, resulting in leakage of an entrapped marker, calcein. The aggregation increased with increasing amount of the lectin; however, the leakage rate was maximum at an optimum concentration of WGA. These results are discussed in terms of the role of sialic acid in the interfacial hydration and charge repulsion which determines the DOPE bilayer stability.
Pinnaduwage
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P
Department of Biochemistry, University of Tennessee, Knoxville 37996-0840.
Huang
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L
D002240:Carbohydrate Sequence; D002846:Chromatography, Affinity; D005452:Fluoresceins; D006021:Glycophorins / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007202:Indicators and Reagents; D008051:Lipid Bilayers; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009844:Oligosaccharides / Q000378:metabolism*; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D011485:Protein Binding; D012795:Sialoglycoproteins / Q000378:metabolism*
D005452:Fluoresceins; D006021:Glycophorins; D007202:Indicators and Reagents; D008051:Lipid Bilayers; D009844:Oligosaccharides; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D012795:Sialoglycoproteins; C020888:1,2-dielaidoylphosphatidylethanolamine; C017251:1,2-oleoylphosphatidylcholine; C007740:fluorexon