Pyrene-labeled gangliosides: micelle formation in aqueous solution, lateral diffusion, and thermotropic behavior in...

By use of the excimer technique, the formation in aqueous solution of pyrene-labeled ganglioside micelles and their lateral diffusion and distribution in phosphatidylcholine membranes were investigated. For these studies 12-(1-pyrenyl)dodecanoic acid was covalently attached to the ceramide part of lysogangliosides GM1, GM2, GM3, GD1a, and GD1b. The 12-(1-pyrenyl)dodecanoic acid substitute of phosphatidylcholine was used for comparison. All pyrene-labeled gangliosides were present in aqueous solution in a predominantly micellar form down to 2 X 10(-8) M, which is the technical limit of this method. The tendency to aggregate is highest for PyGD1a and PyGD1b. In fluid dipalmitoylphosphatidylcholine bilayers the excimer-to-monomer fluorescence intensity ratio of pyrene-labeled gangliosides PyGM1, PyGM2, PyGM3, PyGD1a, and PyGD1b increases linearly with ganglioside concentration. The calculated diffusion coefficients for gangliosides are comparable to 1.6 X 10(-7) cm2/s, which is the diffusion coefficient of pyrene-labeled phosphatidylcholine [Galla, H.-J., & Hartmann, W. (1980) Chem. Phys. Lipids 27, 199-219]. In comparison to phosphatidylcholine, the diffusion of monosialogangliosides is slightly increased, with that diffusion of disialogangliosides being slightly decreased. Ca2+ ions up to 200 mM do not affect ganglioside diffusion significantly. The shape of the lipid phase transition curves obtained by the excimer technique yields information on the lateral distribution of the tested probe molecules. Pyrene-labeled phosphatidylcholine was taken as reference for a system with complete miscibility but nonideal mixing. 1-Acyl-2-[10-(1-pyrenyl)decanoyl]-sn-glycero-3-phosphocholine (PyPC) is known to be randomly distributed in the gel and in the fluid-crystalline lipid phase of dipalmitoylphosphatidylcholine bilayer membranes. It distributes preferentially into the fluid phase in the phase-transition region. In comparison, PyPC in dimyristoylphosphatidylcholine membranes is an example of a system with nearly ideal mixing [Hresko, R. C., Sugar, J. P., Barenholz, Y., & Thompson, T. E. (1986) Biochemistry 25, 3813-3828]. Phase-transition curves of pyrene-labeled gangliosides exemplify a nearly ideal mixing system with PyGD1a or PyGD1b producing best effects. The monosialogangliosides, however, exhibit less ideality of mixing, the deviation from an ideal mixing behavior increasing with decreasing number of both neutral sugar residues and sialic acid groups. Addition of Ca2+ triggers a tightening of the phosphatidylcholine bilayer and thus induces a change in the lateral distribution of the gangliosides at the phase transition.(ABSTRACT TRUNCATED AT 400 WORDS)