Spontaneous Transfer of Ganglioside GMi from Its Micelles to Lipid Vesicles of Differing Size

Rhoderick E Brown; Kristi J. Hyland

doi:10.1021/bi00158a024

Spontaneous Transfer of Ganglioside GMi from Its Micelles to Lipid Vesicles of Differing Size

Rhoderick E Brown, Kristi J. Hyland

Hormel Institute

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

The spontaneous incorporation of II³-N-acetylneuraminosylgangliotetraosylceramide (GM₁) from its micelles into phospholipid bilayer vesicles has been investigated to determine whether curvature-induced changes in membrane lipid packing influence ganglioside uptake. Use of conventional liquid chromatography in conjunction with technically-improved molecular sieve gels permits ganglioside micelles to be separated from phospholipid vesicles of different average size including vesicles with diameters smaller than 40 nm and, thus, allows detailed study of native ganglioside GM₁ incorporation into model membranes under conditions where complicating processes like fusion are readily detected if present. At 45 °C, the spontaneous transfer rate of GMi from its micelles to small unilamellar vesicles (SUVs) comprised of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) is at least 3-fold faster than that to similar composition large unilamellar vesicles (LUVs) prepared by octyl glucoside dialysis. Careful analysis of ganglioside GM₁ distribution among vesicle populations of differing average size reveals that GM₁ preferentially incorporates into the smaller vesicles of certain populations. This behavior is observed in SUVs as well as in LUV-SUV mixtures and actually serves as a sensitive indicator for the presence of trace quantities of SUVs in various LUV preparations. Analysis of the results shows that both differences in the diffusional collision frequency between GM₁ monomers and either SUVs or LUVs and curvature-induced changes in the interfacial lipid packing in either SUVs or LUVs can dramatically influence spontaneous ganglioside uptake. The results have important implications for physiological studies involving sphingolipid-mediated cell signaling events as well as diagnostic paradigms of lysosomal sphingolipid storage diseases in which radiolabeled or fluorescent gangliosides are incorporated into cultured cells by coincubation.

Original language	English (US)
Pages (from-to)	10602-10609
Number of pages	8
Journal	Biochemistry
Volume	31
Issue number	43
DOIs	https://doi.org/10.1021/bi00158a024
State	Published - Feb 1 1992

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title = "Spontaneous Transfer of Ganglioside GMi from Its Micelles to Lipid Vesicles of Differing Size",

abstract = "The spontaneous incorporation of II3-N-acetylneuraminosylgangliotetraosylceramide (GM1) from its micelles into phospholipid bilayer vesicles has been investigated to determine whether curvature-induced changes in membrane lipid packing influence ganglioside uptake. Use of conventional liquid chromatography in conjunction with technically-improved molecular sieve gels permits ganglioside micelles to be separated from phospholipid vesicles of different average size including vesicles with diameters smaller than 40 nm and, thus, allows detailed study of native ganglioside GM1 incorporation into model membranes under conditions where complicating processes like fusion are readily detected if present. At 45 °C, the spontaneous transfer rate of GMi from its micelles to small unilamellar vesicles (SUVs) comprised of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) is at least 3-fold faster than that to similar composition large unilamellar vesicles (LUVs) prepared by octyl glucoside dialysis. Careful analysis of ganglioside GM1 distribution among vesicle populations of differing average size reveals that GM1 preferentially incorporates into the smaller vesicles of certain populations. This behavior is observed in SUVs as well as in LUV-SUV mixtures and actually serves as a sensitive indicator for the presence of trace quantities of SUVs in various LUV preparations. Analysis of the results shows that both differences in the diffusional collision frequency between GM1 monomers and either SUVs or LUVs and curvature-induced changes in the interfacial lipid packing in either SUVs or LUVs can dramatically influence spontaneous ganglioside uptake. The results have important implications for physiological studies involving sphingolipid-mediated cell signaling events as well as diagnostic paradigms of lysosomal sphingolipid storage diseases in which radiolabeled or fluorescent gangliosides are incorporated into cultured cells by coincubation.",

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N2 - The spontaneous incorporation of II3-N-acetylneuraminosylgangliotetraosylceramide (GM1) from its micelles into phospholipid bilayer vesicles has been investigated to determine whether curvature-induced changes in membrane lipid packing influence ganglioside uptake. Use of conventional liquid chromatography in conjunction with technically-improved molecular sieve gels permits ganglioside micelles to be separated from phospholipid vesicles of different average size including vesicles with diameters smaller than 40 nm and, thus, allows detailed study of native ganglioside GM1 incorporation into model membranes under conditions where complicating processes like fusion are readily detected if present. At 45 °C, the spontaneous transfer rate of GMi from its micelles to small unilamellar vesicles (SUVs) comprised of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) is at least 3-fold faster than that to similar composition large unilamellar vesicles (LUVs) prepared by octyl glucoside dialysis. Careful analysis of ganglioside GM1 distribution among vesicle populations of differing average size reveals that GM1 preferentially incorporates into the smaller vesicles of certain populations. This behavior is observed in SUVs as well as in LUV-SUV mixtures and actually serves as a sensitive indicator for the presence of trace quantities of SUVs in various LUV preparations. Analysis of the results shows that both differences in the diffusional collision frequency between GM1 monomers and either SUVs or LUVs and curvature-induced changes in the interfacial lipid packing in either SUVs or LUVs can dramatically influence spontaneous ganglioside uptake. The results have important implications for physiological studies involving sphingolipid-mediated cell signaling events as well as diagnostic paradigms of lysosomal sphingolipid storage diseases in which radiolabeled or fluorescent gangliosides are incorporated into cultured cells by coincubation.

AB - The spontaneous incorporation of II3-N-acetylneuraminosylgangliotetraosylceramide (GM1) from its micelles into phospholipid bilayer vesicles has been investigated to determine whether curvature-induced changes in membrane lipid packing influence ganglioside uptake. Use of conventional liquid chromatography in conjunction with technically-improved molecular sieve gels permits ganglioside micelles to be separated from phospholipid vesicles of different average size including vesicles with diameters smaller than 40 nm and, thus, allows detailed study of native ganglioside GM1 incorporation into model membranes under conditions where complicating processes like fusion are readily detected if present. At 45 °C, the spontaneous transfer rate of GMi from its micelles to small unilamellar vesicles (SUVs) comprised of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) is at least 3-fold faster than that to similar composition large unilamellar vesicles (LUVs) prepared by octyl glucoside dialysis. Careful analysis of ganglioside GM1 distribution among vesicle populations of differing average size reveals that GM1 preferentially incorporates into the smaller vesicles of certain populations. This behavior is observed in SUVs as well as in LUV-SUV mixtures and actually serves as a sensitive indicator for the presence of trace quantities of SUVs in various LUV preparations. Analysis of the results shows that both differences in the diffusional collision frequency between GM1 monomers and either SUVs or LUVs and curvature-induced changes in the interfacial lipid packing in either SUVs or LUVs can dramatically influence spontaneous ganglioside uptake. The results have important implications for physiological studies involving sphingolipid-mediated cell signaling events as well as diagnostic paradigms of lysosomal sphingolipid storage diseases in which radiolabeled or fluorescent gangliosides are incorporated into cultured cells by coincubation.

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Spontaneous Transfer of Ganglioside GMi from Its Micelles to Lipid Vesicles of Differing Size

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