The Influence of Membrane Lipids in Staphylococcus aureus Gamma-Hemolysins Pore Formation

Abstract

The natural target of Staphylococcus aureus bicomponent γ-hemolysins are leucocyte cell membranes. Because a proteinaceous receptor has not been found yet, we checked for the importance of the different membrane lipid compositions by measuring the activity of the toxin on several pure lipid model membranes. We investigated the effect of membrane thickness, fluidity, and presence of nonbilayer lipids and found that the toxin pore-forming ability increased in the presence of phosphocholines with short saturated acyl chains or with unsaturated chains even though not short. An increase of activity was also evident in the presence of cone-shaped lipids like phosphatidylethanolamine or diphytanoylphosphatidylcholine, whereas cylindrical lipids, like sphingomyelin, did not favor the activity. All these results suggest that γ-hemolysins could bind to the bilayer only if the phosphatidylcholine (PC) head is freely accessible. This condition is satisfied by the concurrent presence of cholesterol and certain lipids, as highlighted by the so-called umbrella model (J. Huang and G. W. Feigenson, Biophys J 76:2142–2157, 1999). According to this model, cholesterol could help to a better exposition of PC head groups only if acyl chains are short or unsaturated. In fact, phosphatidylcholines with more than 13 carbon atoms acyl chains can cover cholesterol molecules; in this way, PC head groups pack tightly, rendering them inaccessible to the toxin, which thus shows a reduced pore-forming ability.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Abbreviations

HlgA/HlgB:

Staphylococcus aureus γ-hemolysins A and B

Cho:

Cholesterol

PC:

Phosphatidylcholine

DePC:

DiElaidoyl PC

DMoPC:

DiMyristoleoyl PC

DPoPC:

DiPalmitoleoyl PC

DOPC:

DiOleoyl PC

DPhPC:

DiPhytanoyl PC

DPPC:

Dipalmitoyl PC

PE:

Egg Phosphatidyl-ethanolamine

POPC:

Palmitoyl Oleoyl PC

SM:

Sphingomyelin

Tm :

Main transition temperature

References

  1. Ali MR, Cheng KH, Huang J (2008) Assess the nature of cholesterol–lipid interactions through the chemical potential of cholesterol in phosphatidylcholine bilayers. Proc Natl Acad Sci USA 104:5372–5377

    Article  CAS  Google Scholar 

  2. Bakan E, Yildirim A, Kurtul N, Polat MF, Dursun H, Cayir K (2006) Effects of type 2 diabetes mellitus on plasma fatty acid composition and cholesterol content of erythrocyte and leukocyte membranes. Acta Diabetol 43:109–113

    PubMed  Article  CAS  Google Scholar 

  3. Brassard P, Larbi A, Grenier A, Frisch F, Fortin C, Carpentier AC, Fulop T (2007) Modulation of T-cell signalling by non-esterified fatty acids. Prostaglandins Leukot Essent Fatty Acids 77:337–343

    PubMed  Article  CAS  Google Scholar 

  4. Chiu SW, Jakobsson E, Mashl RJ, Scott HL (2002) Cholesterol-induced modifications in lipid bilayers: a simulation study. Biophys J 83:1842–1853

    PubMed  CAS  Google Scholar 

  5. Crowe JH, Tablin F, Tsvetkova N, Oliver AE, Walker N, Crowe LM (1999) Are lipid phase transitions responsible for chilling damage in human platelets? Cryobiology 38:180–191

    PubMed  Article  CAS  Google Scholar 

  6. Erman F, Aydin S, Demir Y, Akcay F, Bakan E (2006) Determination of saturated and unsaturated Fatty acids amount in leukocyte membranes from subjects fed with solid and fluid oils. J Biochem Mol Biol 39:516–521

    PubMed  CAS  Google Scholar 

  7. Ferreras M, Hoeper F, Dalla Serra M, Colin DA, Prévost G, Menestrina G (1998) The interaction of Staphylococcus aureus bi-component gamma hemolysins and leucocidins with cells and model membranes. Biochim Biophys Acta 1414:108–126

    PubMed  Article  CAS  Google Scholar 

  8. Galdiero S, Gouaux E (2004) High resolution crystallographic studies of alpha-hemolysin-phospholipid complexes define heptamer-lipid head group interactions: implication for understanding protein-lipid interactions. Protein Sci 13:1503–1511

    PubMed  Article  CAS  Google Scholar 

  9. Gauduchon V, Werner S, Prévost G, Monteil H, Colin DA (2001) Flow cytometric determination of Panton-Valentine leucocidin S component binding. Infect Immun 69:2390–2395

    PubMed  Article  CAS  Google Scholar 

  10. Gottfried EL (1971) Lipid patterns in human leukocytes maintained in long-term culture. J Lipid Res 12:531–537

    PubMed  CAS  Google Scholar 

  11. Guillet V, Roblin P, Werner S, Coraiola M, Menestrina G, Monteil H, Prévost G, Mourey L (2004) Crystal structure of leucotoxin S component: new insight into the staphylococcal beta-barrel pore-forming toxins. J Biol Chem 279:41028–41037

    PubMed  Article  CAS  Google Scholar 

  12. Huang J, Feigenson GW (1999) A microscopic interaction model of maximum solubility of cholesterol in lipid bilayers. Biophys J 76:2142–2157

    PubMed  CAS  Google Scholar 

  13. Huang J, Buboltz JT, Feigenson GW (1999) Maximum solubility of cholesterol in phosphatidylcholine and phosphatidylethanolamine bilayers. Biochim Biophys Acta 1417:89–100

    PubMed  Article  CAS  Google Scholar 

  14. Ikigai H, Nakae T (1984) The rate assay of alpha-toxin assembly in membrane. FEMS Microbiol Lett 24:319–322

    Article  CAS  Google Scholar 

  15. Joubert O, Voegelin J, Guillet V, Tranier S, Werner S, Colin DA, Dalla Serra M, Keller D, Monteil H, Mourey L, Prévost G (2007) Distinction between pore assembly by staphylococcal α-toxin versus leucotoxins. J Biomed Biotechnol ID 25935:1–13

    Article  CAS  Google Scholar 

  16. Kaneko J, Kamio Y (2004) Bacterial two-component and hetero-heptameric pore-forming cytolytic toxins: structures, pore-forming mechanism, and organization of the genes. Biosci Biotechnol Biochem 68:981–1003

    PubMed  Article  CAS  Google Scholar 

  17. Labrouche S, Freyburger G, Gin H, Boisseau MR, Cassagne C (1996) Changes in phospholipid composition of blood cell membranes (erythrocyte, platelet, and polymorphonuclear) in different types of diabetes—clinical and biological correlations. Metabolism 45:57–71

    PubMed  Article  CAS  Google Scholar 

  18. Lalazar G, Ben Ya’acov A, Lador A, Livovsky DM, Pappo O, Preston S, Hareati M, Ilan Y (2008) Modulation of intracellular machinery by beta-glycolipids is associated with alteration of NKT lipid rafts and amelioration of concanavalin-induced hepatitis. Mol Immunol 45:3517–3525

    PubMed  Article  CAS  Google Scholar 

  19. Larbi A, Dupuis G, Khalil A, Douziech N, Fortin C, Fulop T Jr (2006) Differential role of lipid rafts in the functions of CD4+ and CD8+ human T lymphocytes with aging. Cell Signal 18:1017–1030

    PubMed  Article  CAS  Google Scholar 

  20. Lawrence JC, Saslowsky DE, Edwardson JM, Henderson RM (2003) Real-time analysis of the effects of cholesterol on lipid raft behavior using atomic force microscopy. Biophys J 84:1827–1832

    PubMed  CAS  Google Scholar 

  21. Leidl K, Liebisch G, Richter D, Schmitz G (2008) Mass spectrometric analysis of lipid species of human circulating blood cells. Biochim Biophys Acta 1781:655–664

    PubMed  CAS  Google Scholar 

  22. Lewis BA, Engelman DM (1983) Lipid bilayer thickness varies linearly with acyl chain length in fluid phosphatidylcholine vesicles. J Mol Biol 166:211–217

    Google Scholar 

  23. MacDonald RC, MacDonald RI, Menco BP, Takeshita K, Subbarao NK, Hu LR (1991) Small-volume extrusion apparatus for preparation of large, unilamellar vesicles. Biochim Biophys Acta 1061:297–303

    PubMed  Article  CAS  Google Scholar 

  24. McIntosh TJ, Vidal A, Simon SA (2003) Sorting of lipids and transmembrane peptides between detergent-soluble bilayers and detergent-resistant rafts. Biophys J 85:1656–1666

    PubMed  CAS  Google Scholar 

  25. Menestrina G, Dalla Serra M, Prévost G (2001) Mode of action of beta-barrel pore-forming toxins of the staphylococcal gamma-hemolysin family. Toxicon 39:1661–1672

    PubMed  Article  CAS  Google Scholar 

  26. Menestrina G, Dalla Serra M, Comai M, Coraiola M, Viero G, Werner S, Colin DA, Monteil H, Prévost G (2003) Ion channels and bacterial infection: the case of beta-barrel pore-forming protein toxins of Staphylococcus aureus. FEBS Lett 552:54–60

    PubMed  Article  CAS  Google Scholar 

  27. Meunier O, Falkenrodt A, Monteil H, Colin DA (1995) Application of flow cytometry in toxinology: pathophysiology of human polymorphonuclear leukocytes damaged by a pore-forming toxin from Staphylococcus aureus. Cytometry 21:241–247

    PubMed  Article  CAS  Google Scholar 

  28. Meunier O, Ferreras M, Supersac G, Hoeper F, Baba-Moussa L, Monteil H, Colin DA, Menestrina G, Prévost G (1997) A predicted beta-sheet from class S components of staphylococcal gamma-hemolysins is essential for the secondary interaction of the class F component. Biochim Biophys Acta 1326:275–286

    Google Scholar 

  29. Nishiyama A, Kaneko J, Harata M, Kamio Y (2006) Assembly of staphylococcal leukocidin into a pore-forming oligomer on detergent-resistant membrane microdomains, lipid rafts, in human polymorphonuclear leukocytes. Biosci Biotechnol Biochem 70:1300–1307

    PubMed  Article  CAS  Google Scholar 

  30. Olson R, Nariya H, Yokota K, Kamio Y, Gouaux JE (1999) Crystal structure of staphylococcal LukF delineates conformational changes accompanying formation of transmembrane channel. Nat Struct Biol 6:134–140

    PubMed  Article  CAS  Google Scholar 

  31. Orr G, Hu D, Ozcelik S, Opresko LK, Wiley HS, Colson SD (2005) Cholesterol dictates the freedom of EGF receptors and HER2 in the plane of the membrane. Biophys J 89:1362–1373

    PubMed  Article  CAS  Google Scholar 

  32. Pandit SA, Bostick D, Berkowitz ML (2004a) Complexation of phosphatidylcholine lipids with cholesterol. Biophys J 86:1345–1356

    PubMed  CAS  Google Scholar 

  33. Pandit SA, Jakobsson E, Scott HL (2004b) Simulation of the early stages of nano-domain formation in mixed bilayers of sphingomyelin, cholesterol, and dioleylphosphatidylcholine. Biophys J 87:3312–3322

    PubMed  Article  CAS  Google Scholar 

  34. Parker A, Miles K, Cheng KH, Huang J (2004) Lateral distribution of cholesterol in dioleoylphosphatidylcholine lipid bilayers: cholesterol–phospholipid interactions at high cholesterol limit. Biophys J 86:1532–1544

    PubMed  CAS  Article  Google Scholar 

  35. Prévost G, Menestrina G, Colin DA, Werner S, Bronner S, Dalla Serra M, Baba Moussa L, Coraiola M, Gravet A, Monteil H (2003) Staphylococcal bicomponent leucotoxins, mechanism of action, impact on cells and contribution to virulence. In: Menestrina G (ed) Pore-forming peptides and protein toxins. Taylor and Francis, London, pp 3–26

    Google Scholar 

  36. Prévost G, Mourey L, Colin DA, Monteil H, Dalla Serra M, Menestrina G (2005) Alpha-helix and beta-barrel pore-forming toxins (leucocidins, alpha-, gamma- and delta-cytolysins) of Staphylococcus aureus. In: Alouf JE, Freer JH (eds) The comprehensive sourcebook of bacterial protein toxins. Academic Press, Amsterdam, pp 588–605

    Google Scholar 

  37. Rouquette-Jazdanian AK, Foussat A, Lamy L, Pelassy C, Lagadec P, Breittmayer JP, Aussel C (2005) Cholera toxin B-subunit prevents activation and proliferation of human CD4+ T cells by activation of a neutral sphingomyelinase in lipid rafts. J Immunol 175:5637–5648

    PubMed  CAS  Google Scholar 

  38. Rouzer CA, Ivanova PT, Byrne MO, Milne SB, Marnett LJ, Brown HA (2006) Lipid profiling reveals arachidonate deficiency in RAW264.7 cells: structural and functional implications. Biochemistry 45:14795–14808

    PubMed  Article  CAS  Google Scholar 

  39. Scherfeld D, Kahya N, Schwille P (2003) Lipid dynamics and domain formation in model membranes composed of ternary mixtures of unsaturated and saturated phosphatidylcholines and cholesterol. Biophys J 85:3758–3768

    PubMed  CAS  Google Scholar 

  40. Simons K, Ikonen E (1997) Functional rafts in cell membranes. Nature 387:569–572

    PubMed  Article  CAS  Google Scholar 

  41. Song L, Hobaugh MR, Shustak C, Cheley S, Bayley H, Gouaux JE (1996) Structure of staphylococcal alpha-hemolysin, a heptameric transmembrane pore. Science 274:1859–1866

    PubMed  Article  CAS  Google Scholar 

  42. Tablin F, Oliver AE, Walker NJ, Crowe LM, Crowe JH (1996) Membrane phase transition of intact human platelets: correlation with cold-induced activation. J Cell Physiol 168:305–313

    PubMed  Article  CAS  Google Scholar 

  43. Tejuca M, Dalla Serra M, Ferreras M, Lanio ME, Menestrina G (1996) The mechanism of membrane permeabilisation by sticholysin I, a cytolysin isolated from the venom of the sea anemone Stichodactyla helianthus. Biochemistry 35:14947–14957

    PubMed  Article  CAS  Google Scholar 

  44. Tilley SJ, Saibil HR (2006) The mechanism of pore formation by bacterial toxins. Curr Opin Struct Biol 16:230–236

    PubMed  Article  CAS  Google Scholar 

  45. Tomita T, Watanabe M, Yasuda T (1992a) Effect of fatty acyl domain of phospholipids on the membrane-channel formation of Staphylococcus aureus alpha-toxin in liposome membrane. Biochim Biophys Acta 1104:325–330

    PubMed  Article  CAS  Google Scholar 

  46. Tomita T, Watanabe M, Yasuda T (1992b) Influence of membrane fluidity on the assembly of Staphylococcus aureus alpha-toxin, a channel-forming protein, in liposome membrane. J Biol Chem 267:13391–13397

    PubMed  CAS  Google Scholar 

  47. Tsvetkova NM, Horváth I, Török Z, Wolkers WF, Balogi Z, Shigapova N, Crowe LM, Tablin F, Vierling E, Crowe JH, Vigh L (2002) Small heat-shock proteins regulate membrane lipid polymorphism. Proc Natl Acad Sci USA 99:13504–13509

    PubMed  Article  CAS  Google Scholar 

  48. Valeva A, Hellmann N, Walev I, Strand D, Plate M, Boukhallouk F, Brack A, Hanada K, Decker H, Bhakdi S (2006) Evidence that clustered phosphocholine head groups serve as sites for binding and assembly of an oligomeric protein pore. J Biol Chem 281:26014–26021

    PubMed  Article  CAS  Google Scholar 

  49. van den Brink-van der Laan E, Killian JA, de Kruijff B (2004) Nonbilayer lipids affect peripheral and integral membrane proteins via changes in the lateral pressure profile. Biochim Biophys Acta 1666:275–288

  50. Viero G, Gropuzzo A, Joubert O, Keller D, Prévost G, Dalla Serra M (2008) A molecular pin to study the dynamic of β-barrel formation in pore forming toxins on erythrocytes: a sliding model. Cell Mol Life Sci 65:312–323

    PubMed  Article  CAS  Google Scholar 

  51. Vrljic M, Nishimura SY, Moerner WE, McConnell HM (2005) Cholesterol depletion suppresses the translational diffusion of class II major histocompatibility complex proteins in the plasma membrane. Biophys J 88:334–347

    PubMed  Article  CAS  Google Scholar 

  52. Werner S, Colin DA, Coraiola M, Menestrina G, Monteil H, Prévost G (2002) Retrieving biological activity from LukF-PV mutants combined with different S-components implies compatibility between stem domains of these staphylococcal bi-component leucotoxins. Infect Immun 70:1310–1318

    PubMed  Article  CAS  Google Scholar 

  53. Wolkers WF, Looper SA, Fontanilla RA, Tsvetkova NM, Tablin F, Crowe JH (2003) Temperature dependence of fluid phase endocytosis coincides with membrane properties of pig platelets. Biochim Biophys Acta 1612:154–163

    PubMed  Article  CAS  Google Scholar 

  54. Xu XL, London E (2000) The effect of sterol structure on membrane lipid domains reveals how cholesterol can induce lipid domain formation. Biochemistry 39:843–849

    PubMed  Article  CAS  Google Scholar 

  55. Zitzer A, Bittman R, Verbicky CA, Erukulla RK, Bhakdi S, Weis S, Valeva A, Palmer M (2001) Coupling of cholesterol and cone-shaped lipids in bilayers augments membrane permeabilization by the cholesterol-specific toxins streptolysin O and Vibrio cholerae cytolysin. J Biol Chem 276:14628–14633

    PubMed  Article  CAS  Google Scholar 

Download references

Acknowledgments

This work started under the supervision of Gianfranco Menestrina, and we dedicate this article to his memory. The present work was supported by the Consiglio Nazionale delle Ricerche (CNR), the Istituto Trentino di Cultura (ITC), the Provincia Autonoma di Trento (PAT) Fondo Progetti (Projects StaWars and Plugs), and EA-3432 from the “Institut de Bactériologie de la Faculté de Médecine,” Strasbourg. It was also partially sponsored by Fondazione Cariverona, Bando 2003.

Author information

Affiliations

Authors

Corresponding author

Correspondence to C. Potrich.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Potrich, C., Bastiani, H., Colin, D.A. et al. The Influence of Membrane Lipids in Staphylococcus aureus Gamma-Hemolysins Pore Formation. J Membrane Biol 227, 13 (2009). https://doi.org/10.1007/s00232-008-9140-6

Download citation

Keywords

  • γ-Hemolysins
  • Cone-shaped lipids
  • Umbrella model
  • Phosphocholine head groups