Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1980 May;77(5):2362–2366. doi: 10.1073/pnas.77.5.2362

Lipid-protein interaction in the glycophorin-dipalmitoylphosphatidylcholine system: Raman spectroscopic investigation

Theodore Taraschi 1, Richard Mendelsohn 1,*
PMCID: PMC349397  PMID: 16592811

Abstract

Raman spectra have been recorded as a function of temperature for lipid-protein complexes of glycophorin isolated from erythrocyte membranes reconstituted with dipalmitoylphosphatidylcholine (DPPC) and its chain perdeuterated analogue ([2H62]DPPC). The conformation of the phospholipid hydrocarbon chains in the vicinity of protein is drastically altered from that in pure lipid dispersions. Analysis of the chain C-2H stretching vibrations for complexes of [2H62]-DPPC-glycophorin shows that at lipid:protein mole ratios of 125:1, a broad melting event occurs that is not observable by calorimetric techniques. The midpoint occurs at temperatures about 15°C below that of the gel/liquid crystal phase transition for [2H62]DPPC in multilamellar dispersions. The same number of gauche rotamers form in the phospholipid hydrocarbon chains during the melting process as in the phase transition of the unperturbed molecule. Analysis of the C-H stretching region of the Raman spectrum in DPPC-glycophorin complexes indicates that lateral interactions between phospholipid chains in the complex are reduced so that interchain vibrational coupling is minimized. The observed differences between the Raman melting curves and the calorimetric endothermic transitions arise because different populations of phospholipid molecules are sampled in the two experiments. The advantages of Raman spectroscopy for the study of lipid-protein interaction are demonstrated in the current work. Implications for the structure of the lipid in the immediate vicinity of membrane protein are discussed.

Keywords: lipid phase transitions, vibrational spectroscopy, deuterated lipids

Full text

PDF
2362

Selected References

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

  1. Bennett J. P., McGill K. A., Warren G. B. Transbilayer disposition of the phospholipid annulus surrounding a calcium transport protein. Nature. 1978 Aug 24;274(5673):823–825. doi: 10.1038/274823a0. [DOI] [PubMed] [Google Scholar]
  2. Brûlet P., McConnell H. M. Protein-lipid interactions glycophorin and dipalmitolyphosphatidylcholine. Biochem Biophys Res Commun. 1976 Jan 26;68(2):363–368. doi: 10.1016/0006-291x(76)91153-0. [DOI] [PubMed] [Google Scholar]
  3. DODGE J. T., MITCHELL C., HANAHAN D. J. The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes. Arch Biochem Biophys. 1963 Jan;100:119–130. doi: 10.1016/0003-9861(63)90042-0. [DOI] [PubMed] [Google Scholar]
  4. Dahlquist F. W., Muchmore D. C., Davis J. H., Bloom M. Deuterium magnetic resonance studies of the interaction of lipids with membrane proteins. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5435–5439. doi: 10.1073/pnas.74.12.5435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Davis D. G., Inesi G., Gulik-Krzywicki T. Lipid molecular motion and enzyme activity in sarcoplasmic reticulum membrane. Biochemistry. 1976 Mar 23;15(6):1271–1276. doi: 10.1021/bi00651a016. [DOI] [PubMed] [Google Scholar]
  6. Gaber B. P., Peticolas W. L. On the quantitative interpretation of biomembrane structure by Raman spectroscopy. Biochim Biophys Acta. 1977 Mar 1;465(2):260–274. doi: 10.1016/0005-2736(77)90078-5. [DOI] [PubMed] [Google Scholar]
  7. Gennis R. B. Protein-lipid interactions. Annu Rev Biophys Bioeng. 1977;6:195–238. doi: 10.1146/annurev.bb.06.060177.001211. [DOI] [PubMed] [Google Scholar]
  8. Gerritsen W. J., van Zoelen E. J., Verkleij A. J., de Kruijff B., van Deenen L. L. A 13C NMR method for determination of the transbilayer distribution of phosphatidylcholine in large, unilamellar, protein-free and protein-containing vesicles. Biochim Biophys Acta. 1979 Mar 8;551(2):248–259. doi: 10.1016/0005-2736(89)90003-5. [DOI] [PubMed] [Google Scholar]
  9. Grant C. W., McConnell H. M. Glycophorin in lipid bilayers. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4653–4657. doi: 10.1073/pnas.71.12.4653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jost P. C., Griffith O. H., Capaldi R. A., Vanderkooi G. Evidence for boundary lipid in membranes. Proc Natl Acad Sci U S A. 1973 Feb;70(2):480–484. doi: 10.1073/pnas.70.2.480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  12. Lippert J. L., Gorczyca L. E., Meiklejohn G. A laser Raman spectroscopic investigation of phospholipid and protein configurations in hemoglobin-free erythrocyte ghosts. Biochim Biophys Acta. 1975 Feb 28;382(1):51–57. doi: 10.1016/0005-2736(75)90371-5. [DOI] [PubMed] [Google Scholar]
  13. Mabrey S., Sturtevant J. M. Investigation of phase transitions of lipids and lipid mixtures by sensitivity differential scanning calorimetry. Proc Natl Acad Sci U S A. 1976 Nov;73(11):3862–3866. doi: 10.1073/pnas.73.11.3862. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Marchesi V. T., Andrews E. P. Glycoproteins: isolation from cellmembranes with lithium diiodosalicylate. Science. 1971 Dec 17;174(4015):1247–1248. doi: 10.1126/science.174.4015.1247. [DOI] [PubMed] [Google Scholar]
  15. Marchesi V. T., Furthmayr H., Tomita M. The red cell membrane. Annu Rev Biochem. 1976;45:667–698. doi: 10.1146/annurev.bi.45.070176.003315. [DOI] [PubMed] [Google Scholar]
  16. Mendelsohn R., Maisano J. Use of deuterated phospholipids in Raman spectroscopic studies of membrane structure. I. Multilayers of dimyristoyl phosphatidylcholine (and its -d54 derivative) with distearoyl phosphatidylcholine. Biochim Biophys Acta. 1978 Jan 19;506(2):192–201. doi: 10.1016/0005-2736(78)90390-5. [DOI] [PubMed] [Google Scholar]
  17. Mendelsohn R., Taraschi T. Deuterated phospholipids as Raman spectroscopic probes of membrane structure: dipalmitoylphosphatidylcholine-dipalmitoylphosphatidylethanolamine multilayers. Biochemistry. 1978 Sep 19;17(19):3944–3949. doi: 10.1021/bi00612a010. [DOI] [PubMed] [Google Scholar]
  18. Oldfield E., Gilmore R., Glaser M., Gutowsky H. S., Hshung J. C., Kang S. Y., King T. E., Meadows M., Rice D. Deuterium nuclear magnetic resonance investigation of the effects of proteins and polypeptides on hydrocarbon chain order in model membrane systems. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4657–4660. doi: 10.1073/pnas.75.10.4657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Petersen N. O., Kroon P. A., Kainoshio M., Chan S. I. Thermal phase transitions in deuterated lecithin bilayers. Chem Phys Lipids. 1975 Aug;14(4):343–349. doi: 10.1016/0009-3084(75)90071-7. [DOI] [PubMed] [Google Scholar]
  20. Rothschild K. J., Andrew J. R., De Grip W. J., Stanley H. E. Opsin structure probed by raman spectroscopy of photoreceptor membranes. Science. 1976 Mar 19;191(4232):1176–1178. doi: 10.1126/science.1257742. [DOI] [PubMed] [Google Scholar]
  21. Warren G. B., Houslay M. D., Metcalfe J. C., Birdsall N. J. Cholesterol is excluded from the phospholipid annulus surrounding an active calcium transport protein. Nature. 1975 Jun 26;255(5511):684–687. doi: 10.1038/255684a0. [DOI] [PubMed] [Google Scholar]
  22. Yellin N., Levin I. W. Hydrocarbon trans-gauche isomerization in phospholipid bilayer gel assemblies. Biochemistry. 1977 Feb 22;16(4):642–647. doi: 10.1021/bi00623a014. [DOI] [PubMed] [Google Scholar]
  23. van Zoelen E. J., Verkleij A. J., Zwaal R. F., van Deenen L. L. Incorporation and asymmetric orientation of glycophorin in reconstituted protein-containing vesicles. Eur J Biochem. 1978 May 16;86(2):539–546. doi: 10.1111/j.1432-1033.1978.tb12337.x. [DOI] [PubMed] [Google Scholar]
  24. van Zoelen E. J., van Dijck P. W., de Kruijff B., Verkleij A. J., van Deenen L. L. Effect of glycophorin incorporation on the physico-chemical properties of phospholipid bilayers. Biochim Biophys Acta. 1978 Dec 4;514(1):9–24. doi: 10.1016/0005-2736(78)90073-1. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES