Skip to main content
NCBI home page
Biophysical Journal logoLink to Biophysical Journal
. 1996 Sep;71(3):1400–1421. doi: 10.1016/S0006-3495(96)79342-6

Oriented 1,2-dimyristoyl-sn-glycero-3-phosphorylcholine/ganglioside membranes: a Fourier transform infrared attenuated total reflection spectroscopic study. Band assignments; orientational, hydrational, and phase behavior; and effects of Ca2+ binding.

E Müller 1, A Giehl 1, G Schwarzmann 1, K Sandhoff 1, A Blume 1
PMCID: PMC1233608  PMID: 8874015

Abstract

Fourier transform infrared (FTIR) attenuated total reflection (ATR) spectroscopy was used to elucidate the hydration behavior and molecular order of phospholipid/ganglioside bilayers. We examined dry and hydrated films of the gangliosides GM1, deacetyl-GM1, lyso-GM1, deacetyllyso-GM1, and GM3 and oriented mixed films of these gangliosides with 1,2-dimyristoyl-sn-glycero-3-phosphorylcholine (DMPC) using polarized light. Analysis of the amide I frequencies reveals that the amide groups are involved in intermolecular interactions via hydrogen bonds of varying strengths. The tilt angle of the acyl chains of the lipids in mixed films was determined as a function of ganglioside structure. Deacetylation of the sialic acid in the headgroup has a stronger influence on the tilt angle than the removal of the ganglioside fatty acid. The phase behavior was examined by FTIR ATR spectroscopy and by differential scanning calorimetry (DSC) measurements on lipid suspensions. At the same molar concentration, lyso-gangliosides have less effect on changes of transition temperature compared to the double-chain analogs. Distinct differences in the amide band shapes were observed between mixtures with lyso-gangliosides and normal double-chain gangliosides. Determined from the dicroic ratio RATR, the orientation of the COO- group in all DMPC/ganglioside mixtures was found to be relatively fixed with respect to the membrane normal. In 4:1 mixtures of DMPC with GM1 and deacetyl-GM1, the binding of Ca2+ leads to a slight decrease in chain tilt in the gel phase, probably caused by a dehydration of the membrane-water interface. In mixtures of DMPC with GM3 and deacetyl-lyso-GM1, a slight increase in chain tilt is observed. The chain tilt in DMPC/lyso-GM1 mixtures is unchanged. Analysis of the COO- band reveals that Ca2+ does not bind to the carboxylate group of the sialic acid of GM1 and deacetyl-GM1, the mixtures in which a decrease in chain tilt was observed. Binding to the sialic acid was only observed for mixtures of DMPC with GM3, lyso-GM1, and deacetyl-lyso-GM1. Ca2+ obviously accumulates at the bilayer-water interface and leads to partial dehydration of the headgroup region in the gel as well as in the liquid-crystalline phase. This can be concluded from the changes in the amide I band shapes. With the exception of DMPC/deacetyl-GM1, the effects on the ester C==O bands are small. The addition of Ca2+ has minor effects on the phase behavior, with the exception of the DMPC/GM1 mixture.

Full text

PDF
1400

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bertoli E., Masserini M., Sonnino S., Ghidoni R., Cestaro B., Tettamanti G. Electron paramagnetic resonance studies on the fluidity and surface dynamics of egg phosphatidylcholine vesicles containing gangliosides. Biochim Biophys Acta. 1981 Oct 2;647(2):196–202. doi: 10.1016/0005-2736(81)90246-7. [DOI] [PubMed] [Google Scholar]
  2. Blume A., Hübner W., Messner G. Fourier transform infrared spectroscopy of 13C = O-labeled phospholipids hydrogen bonding to carbonyl groups. Biochemistry. 1988 Oct 18;27(21):8239–8249. doi: 10.1021/bi00421a038. [DOI] [PubMed] [Google Scholar]
  3. Brandenburg K., Seydel U. Orientation measurements on ordered multibilayers of phospholipids and sphingolipids from synthetic and natural origin by ATR Fourier transform infrared spectroscopy. Z Naturforsch C. 1986 Apr;41(4):453–467. doi: 10.1515/znc-1986-0412. [DOI] [PubMed] [Google Scholar]
  4. Casal H. L., Mantsch H. H., Hauser H. Infrared studies of fully hydrated saturated phosphatidylserine bilayers. Effect of Li+ and Ca2+. Biochemistry. 1987 Jul 14;26(14):4408–4416. doi: 10.1021/bi00388a033. [DOI] [PubMed] [Google Scholar]
  5. Casal H. L., Mantsch H. H., Paltauf F., Hauser H. Infrared and 31P-NMR studies of the effect of Li+ and Ca2+ on phosphatidylserines. Biochim Biophys Acta. 1987 Jun 23;919(3):275–286. doi: 10.1016/0005-2760(87)90267-0. [DOI] [PubMed] [Google Scholar]
  6. Casal H. L., Martin A., Mantsch H. H., Paltauf F., Hauser H. Infrared studies of fully hydrated unsaturated phosphatidylserine bilayers. Effect of Li+ and Ca2+. Biochemistry. 1987 Nov 17;26(23):7395–7401. doi: 10.1021/bi00397a030. [DOI] [PubMed] [Google Scholar]
  7. Cumar F. A., Maggio B., Caputto R. Neurotransmitter movements in nerve endings. Influence of substances that modify the interfacial potential. Biochim Biophys Acta. 1980 Mar 27;597(1):174–182. doi: 10.1016/0005-2736(80)90161-3. [DOI] [PubMed] [Google Scholar]
  8. Fringeli U. P. The structure of lipids and proteins studied by attenuated total reflection (ATR) infrared spectroscopy. II. Oriented layers of a homologous series: phosphatidylethanolamine to phosphatidylcholine. Z Naturforsch C. 1977 Jan-Feb;32(1-2):20–45. doi: 10.1515/znc-1977-1-205. [DOI] [PubMed] [Google Scholar]
  9. Hanai N., Dohi T., Nores G. A., Hakomori S. A novel ganglioside, de-N-acetyl-GM3 (II3NeuNH2LacCer), acting as a strong promoter for epidermal growth factor receptor kinase and as a stimulator for cell growth. J Biol Chem. 1988 May 5;263(13):6296–6301. [PubMed] [Google Scholar]
  10. Hannun Y. A., Bell R. M. Lysosphingolipids inhibit protein kinase C: implications for the sphingolipidoses. Science. 1987 Feb 6;235(4789):670–674. doi: 10.1126/science.3101176. [DOI] [PubMed] [Google Scholar]
  11. Hidari K. I., Irie F., Suzuki M., Kon K., Ando S., Hirabayashi Y. A novel ganglioside with a free amino group in bovine brain. Biochem J. 1993 Nov 15;296(Pt 1):259–263. doi: 10.1042/bj2960259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hinz H. J., Körner O., Nicolau C. Influence of gangliosides GM1 and GD1a on structural and thermotropic properties of sonicated small 1,2-dipalmitoyl-L-alpha-phosphatidylcholine vesicles. Biochim Biophys Acta. 1981 May 20;643(3):557–571. doi: 10.1016/0005-2736(81)90352-7. [DOI] [PubMed] [Google Scholar]
  13. Hübner W., Mantsch H. H. Orientation of specifically 13C=O labeled phosphatidylcholine multilayers from polarized attenuated total reflection FT-IR spectroscopy. Biophys J. 1991 Jun;59(6):1261–1272. doi: 10.1016/S0006-3495(91)82341-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Maggio B., Albert J., Yu R. K. Thermodynamic-geometric correlations for the morphology of self-assembled structures of glycosphingolipids and their mixtures with dipalmitoylphosphatidylcholine. Biochim Biophys Acta. 1988 Nov 22;945(2):145–160. doi: 10.1016/0005-2736(88)90477-4. [DOI] [PubMed] [Google Scholar]
  15. Maggio B., Cumar F. A., Caputto R. Molecular behaviour of glycosphingolipids in interfaces. Possible participation in some properties of nerve membranes. Biochim Biophys Acta. 1981 Dec;650(2-3):69–87. doi: 10.1016/0304-4157(81)90001-0. [DOI] [PubMed] [Google Scholar]
  16. Maggio B. Geometric and thermodynamic restrictions for the self-assembly of glycosphingolipid-phospholipid systems. Biochim Biophys Acta. 1985 May 14;815(2):245–258. doi: 10.1016/0005-2736(85)90295-0. [DOI] [PubMed] [Google Scholar]
  17. Maggio B., Montich G. G., Cumar F. A. Surface topography of sulfatide and gangliosides in unilamellar vesicles of dipalmitoylphosphatidylcholine. Chem Phys Lipids. 1988 Feb;46(2):137–146. doi: 10.1016/0009-3084(88)90124-7. [DOI] [PubMed] [Google Scholar]
  18. Manzi A. E., Sjoberg E. R., Diaz S., Varki A. Biosynthesis and turnover of O-acetyl and N-acetyl groups in the gangliosides of human melanoma cells. J Biol Chem. 1990 Aug 5;265(22):13091–13103. [PubMed] [Google Scholar]
  19. Masserini M., Freire E. Thermotropic characterization of phosphatidylcholine vesicles containing ganglioside GM1 with homogeneous ceramide chain length. Biochemistry. 1986 Mar 11;25(5):1043–1049. doi: 10.1021/bi00353a014. [DOI] [PubMed] [Google Scholar]
  20. McDaniel R., McLaughlin S. The interaction of calcium with gangliosides in bilayer membranes. Biochim Biophys Acta. 1985 Oct 10;819(2):153–160. doi: 10.1016/0005-2736(85)90169-5. [DOI] [PubMed] [Google Scholar]
  21. Mueller E., Blume A. FTIR spectroscopic analysis of the amide and acid bands of ganglioside GM1, in pure form and in mixtures with DMPC. Biochim Biophys Acta. 1993 Feb 23;1146(1):45–51. doi: 10.1016/0005-2736(93)90336-x. [DOI] [PubMed] [Google Scholar]
  22. Okamura E., Umemura J., Takenaka T. Orientation studies of hydrated dipalmitoylphosphatidylcholine multibilayers by polarized FTIR-ATR spectroscopy. Biochim Biophys Acta. 1990 Jun 11;1025(1):94–98. doi: 10.1016/0005-2736(90)90195-t. [DOI] [PubMed] [Google Scholar]
  23. Petersen N. O., Chan S. I. More on the motional state of lipid bilayer membranes: interpretation of order parameters obtained from nuclear magnetic resonance experiments. Biochemistry. 1977 Jun 14;16(12):2657–2667. doi: 10.1021/bi00631a012. [DOI] [PubMed] [Google Scholar]
  24. Probst W., Rösner H., Wiegandt H., Rahmann H. Das Komplexationsvermögen von Gangliosiden für Ca2, I. Einfluss mono- und divalenter Kationen sowie von Acetylcholin. Hoppe Seylers Z Physiol Chem. 1979 Jul;360(7):979–986. [PubMed] [Google Scholar]
  25. Scarsdale J. N., Prestegard J. H., Yu R. K. NMR and computational studies of interactions between remote residues in gangliosides. Biochemistry. 1990 Oct 23;29(42):9843–9855. doi: 10.1021/bi00494a014. [DOI] [PubMed] [Google Scholar]
  26. Schwarzmann G., Sandhoff K. Lysogangliosides: synthesis and use in preparing labeled gangliosides. Methods Enzymol. 1987;138:319–341. doi: 10.1016/0076-6879(87)38028-0. [DOI] [PubMed] [Google Scholar]
  27. Sela B. A., Bach D. Calorimetric studies on the interaction of gangliosides with phospholipids and myelin basic protein. Biochim Biophys Acta. 1984 Apr 11;771(2):177–182. doi: 10.1016/0005-2736(84)90530-3. [DOI] [PubMed] [Google Scholar]
  28. Sharom F. J., Grant C. W. A model for ganglioside behaviour in cell membranes. Biochim Biophys Acta. 1978 Feb 21;507(2):280–293. doi: 10.1016/0005-2736(78)90423-6. [DOI] [PubMed] [Google Scholar]
  29. Sillerud L. O., Prestégard J. H., Yu R. K., Schafer D. E., Konigsberg W. H. Assignment of the 13C nuclear magnetic resonance spectrum of aqueous ganglioside GM1 micelles. Biochemistry. 1978 Jun 27;17(13):2619–2628. doi: 10.1021/bi00606a025. [DOI] [PubMed] [Google Scholar]
  30. Sillerud L. O., Schafer D. E., Yu R. K., Konigsberg W. H. Calorimetric properties of mixtures of ganglioside GM1 and dipalmitoylphosphatidylcholine. J Biol Chem. 1979 Nov 10;254(21):10876–10880. [PubMed] [Google Scholar]
  31. Sonnino S., Cantù L., Corti M., Acquotti D., Venerando B. Aggregative properties of gangliosides in solution. Chem Phys Lipids. 1994 May 6;71(1):21–45. doi: 10.1016/0009-3084(94)02304-2. [DOI] [PubMed] [Google Scholar]
  32. Svennerholm L., Fredman P. A procedure for the quantitative isolation of brain gangliosides. Biochim Biophys Acta. 1980 Jan 18;617(1):97–109. doi: 10.1016/0005-2760(80)90227-1. [DOI] [PubMed] [Google Scholar]
  33. Terzaghi A., Tettamanti G., Masserini M. Interaction of glycosphingolipids and glycoproteins: thermotropic properties of model membranes containing GM1 ganglioside and glycophorin. Biochemistry. 1993 Sep 21;32(37):9722–9725. doi: 10.1021/bi00088a026. [DOI] [PubMed] [Google Scholar]
  34. Uchida T., Nagai Y., Kawasaki Y., Wakayama N. Fluorospectroscopic studies of various ganglioside and ganglioside--lecithin dispersions. Steady-state and time-resolved fluorescence measurements with 1,6-diphenyl-1,3,5-hexatriene. Biochemistry. 1981 Jan 6;20(1):162–169. doi: 10.1021/bi00504a027. [DOI] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES