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Proving Lipid Rafts Exist: Membrane Domains in the Prokaryote Have the Same Properties as Eukaryotic Lipid Rafts


Lipid rafts in eukaryotic cells are sphingolipid and cholesterol-rich, ordered membrane regions that have been postulated to play roles in many membrane functions, including infection. We previously demonstrated the existence of cholesterol-lipid-rich domains in membranes of the prokaryote, B. burgdorferi, the causative agent of Lyme disease [LaRocca et al. (2010) Cell Host & Microbe 8, 331–342]. Here, we show that these prokaryote membrane domains have the hallmarks of eukaryotic lipid rafts, despite lacking sphingolipids. Substitution experiments replacing cholesterol lipids with a set of sterols, ranging from strongly raft-promoting to raft-inhibiting when mixed with eukaryotic sphingolipids, showed that sterols that can support ordered domain formation are both necessary and sufficient for formation of B. burgdorferi membrane domains that can be detected by transmission electron microscopy or in living organisms by Förster resonance energy transfer (FRET). Raft-supporting sterols were also necessary and sufficient for formation of high amounts of detergent resistant membranes from B. burgdorferi. Furthermore, having saturated acyl chains was required for a biotinylated lipid to associate with the cholesterol-lipid-rich domains in B. burgdorferi, another characteristic identical to that of eukaryotic lipid rafts. Sterols supporting ordered domain formation were also necessary and sufficient to maintain B. burgdorferi membrane integrity, and thus critical to the life of the organism. These findings provide compelling evidence for the existence of lipid rafts and show that the same principles of lipid raft formation apply to prokaryotes and eukaryotes despite marked differences in their lipid compositions.


Vyšlo v časopise: Proving Lipid Rafts Exist: Membrane Domains in the Prokaryote Have the Same Properties as Eukaryotic Lipid Rafts. PLoS Pathog 9(5): e32767. doi:10.1371/journal.ppat.1003353
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1003353

Souhrn

Lipid rafts in eukaryotic cells are sphingolipid and cholesterol-rich, ordered membrane regions that have been postulated to play roles in many membrane functions, including infection. We previously demonstrated the existence of cholesterol-lipid-rich domains in membranes of the prokaryote, B. burgdorferi, the causative agent of Lyme disease [LaRocca et al. (2010) Cell Host & Microbe 8, 331–342]. Here, we show that these prokaryote membrane domains have the hallmarks of eukaryotic lipid rafts, despite lacking sphingolipids. Substitution experiments replacing cholesterol lipids with a set of sterols, ranging from strongly raft-promoting to raft-inhibiting when mixed with eukaryotic sphingolipids, showed that sterols that can support ordered domain formation are both necessary and sufficient for formation of B. burgdorferi membrane domains that can be detected by transmission electron microscopy or in living organisms by Förster resonance energy transfer (FRET). Raft-supporting sterols were also necessary and sufficient for formation of high amounts of detergent resistant membranes from B. burgdorferi. Furthermore, having saturated acyl chains was required for a biotinylated lipid to associate with the cholesterol-lipid-rich domains in B. burgdorferi, another characteristic identical to that of eukaryotic lipid rafts. Sterols supporting ordered domain formation were also necessary and sufficient to maintain B. burgdorferi membrane integrity, and thus critical to the life of the organism. These findings provide compelling evidence for the existence of lipid rafts and show that the same principles of lipid raft formation apply to prokaryotes and eukaryotes despite marked differences in their lipid compositions.


Zdroje

1. BenachJL, BoslerEM, HanrahanJP, ColemanJL, HabichtGS, et al. (1983) Spirochetes isolated from the blood of two patients with Lyme disease. N Engl J Med 308: 740–742.

2. BurgdorferW, BarbourAG, HayesSF, BenachJL, GrunwaldtE, et al. (1982) Lyme disease-a tick-borne spirochetosis? Science 216: 1317–1319.

3. JohnsonRC (1977) The spirochetes. Annu Rev Microbiol 31: 89–106.

4. MotalebMA, CorumL, BonoJL, EliasAF, RosaP, et al. (2000) Borrelia burgdorferi periplasmic flagella have both skeletal and motility functions. Proc Natl Acad Sci U S A 97: 10899–10904.

5. LaRoccaTJ, BenachJL (2008) The important and diverse roles of antibodies in the host response to Borrelia infections. Curr Top Microbiol Immunol 319: 63–103.

6. BelisleJT, BrandtME, RadolfJD, NorgardMV (1994) Fatty acids of Treponema pallidum and Borrelia burgdorferi lipoproteins. J Bacteriol 176: 2151–2157.

7. JonesJD, BourellKW, NorgardMV, RadolfJD (1995) Membrane topology of Borrelia burgdorferi and Treponema pallidum lipoproteins. Infect Immun 63: 2424–2434.

8. RadolfJD, GoldbergMS, BourellK, BakerSI, JonesJD, et al. (1995) Characterization of outer membranes isolated from Borrelia burgdorferi, the Lyme disease spirochete. Infect Immun 63: 2154–2163.

9. StubsG, FingerleV, WilskeB, GobelUB, ZahringerU, et al. (2009) Acylated cholesteryl galactosides are specific antigens of Borrelia causing lyme disease and frequently induce antibodies in late stages of disease. J Biol Chem 284: 13326–13334.

10. SchroderNW, SchombelU, HeineH, GobelUB, ZahringerU, et al. (2003) Acylated cholesteryl galactoside as a novel immunogenic motif in Borrelia burgdorferi sensu stricto. J Biol Chem 278: 33645–33653.

11. Ben-MenachemG, Kubler-KielbJ, CoxonB, YergeyA, SchneersonR (2003) A newly discovered cholesteryl galactoside from Borrelia burgdorferi. Proc Natl Acad Sci U S A 100: 7913–7918.

12. WheelerCM, Garcia MoncoJC, BenachJL, GolightlyMG, HabichtGS, et al. (1993) Nonprotein antigens of Borrelia burgdorferi. J Infect Dis 167: 665–674.

13. HaqueM, HiraiY, YokotaK, OgumaK (1995) Steryl glycosides: a characteristic feature of the Helicobacter spp.? J Bacteriol 177: 5334–5337.

14. HiraiY, HaqueM, YoshidaT, YokotaK, YasudaT, et al. (1995) Unique cholesteryl glucosides in Helicobacter pylori: composition and structural analysis. J Bacteriol 177: 5327–5333.

15. LinM, RikihisaY (2003) Ehrlichia chaffeensis and Anaplasma phagocytophilum lack genes for lipid A biosynthesis and incorporate cholesterol for their survival. Infect Immun 71: 5324–5331.

16. SmithPF (1971) Biosynthesis of cholesteryl glucoside by Mycoplasma gallinarum. J Bacteriol 108: 986–991.

17. TrottDJ, AltDP, ZuernerRL, WannemuehlerMJ, StantonTB (2001) The search for Brachyspira outer membrane proteins that interact with the host. Anim Health Res Rev 2: 19–30.

18. SimonsK, van MeerG (1988) Lipid sorting in epithelial cells. Biochemistry 27: 6197–6202.

19. BrownDA, LondonE (2000) Structure and function of sphingolipid- and cholesterol-rich membrane rafts. J Biol Chem 275: 17221–17224.

20. LondonE (2002) Insights into lipid raft structure and formation from experiments in model membranes. Curr Opin Struct Biol 12: 480–486.

21. SchroederR, LondonE, BrownD (1994) Interactions between saturated acyl chains confer detergent resistance on lipids and glycosylphosphatidylinositol (GPI)-anchored proteins: GPI-anchored proteins in liposomes and cells show similar behavior. Proc Natl Acad Sci U S A 91: 12130–12134.

22. AhmedSN, BrownDA, LondonE (1997) On the origin of sphingolipid/cholesterol-rich detergent-insoluble cell membranes: physiological concentrations of cholesterol and sphingolipid induce formation of a detergent-insoluble, liquid-ordered lipid phase in model membranes. Biochemistry 36: 10944–10953.

23. SimonsK, IkonenE (1997) Functional rafts in cell membranes. Nature 387: 569–572.

24. LondonE (2005) How principles of domain formation in model membranes may explain ambiguities concerning lipid raft formation in cells. Biochim Biophys Acta 1746: 203–220.

25. LaRoccaTJ, CrowleyJT, CusackBJ, PathakP, BenachJ, et al. (2010) Cholesterol lipids of Borrelia burgdorferi form lipid rafts and are required for the bactericidal activity of a complement-independent antibody. Cell Host Microbe 8: 331–342.

26. XuX, LondonE (2000) The effect of sterol structure on membrane lipid domains reveals how cholesterol can induce lipid domain formation. Biochemistry 39: 843–849.

27. XuX, BittmanR, DuportailG, HeisslerD, VilchezeC, et al. (2001) Effect of the structure of natural sterols and sphingolipids on the formation of ordered sphingolipid/sterol domains (rafts). Comparison of cholesterol to plant, fungal, and disease-associated sterols and comparison of sphingomyelin, cerebrosides, and ceramide. J Biol Chem 276: 33540–33546.

28. WangJ, Megha, LondonE (2004) Relationship between sterol/steroid structure and participation in ordered lipid domains (lipid rafts): implications for lipid raft structure and function. Biochemistry 43: 1010–1018.

29. Megha, BakhtO, LondonE (2006) Cholesterol precursors stabilize ordinary and ceramide-rich ordered lipid domains (lipid rafts) to different degrees. Implications for the Bloch hypothesis and sterol biosynthesis disorders. J Biol Chem 281: 21903–21913.

30. Garcia MoncoJC, WheelerCM, BenachJL, FurieRA, LukehartSA, et al. (1993) Reactivity of neuroborreliosis patients (Lyme disease) to cardiolipin and gangliosides. J Neurol Sci 117: 206–214.

31. Garcia-MoncoJC, SeidmanRJ, BenachJL (1995) Experimental immunization with Borrelia burgdorferi induces development of antibodies to gangliosides. Infect Immun 63: 4130–4137.

32. KiskowskiMA, HancockJF, KenworthyAK (2009) On the use of Ripley's K-function and its derivatives to analyze domain size. Biophys J 97: 1095–1103.

33. BurandaT, WuY, PerezD, ChigaevA, SklarLA (2010) Real-time partitioning of octadecyl rhodamine B into bead-supported lipid bilayer membranes revealing quantitative differences in saturable binding sites in DOPC and 1∶1∶1 DOPC/SM/cholesterol membranes. J Phys Chem B 114: 1336–1349.

34. LouraLM, FedorovA, PrietoM (2001) Exclusion of a cholesterol analog from the cholesterol-rich phase in model membranes. Biochim Biophys Acta 1511: 236–243.

35. BakhtO, PathakP, LondonE (2007) Effect of the structure of lipids favoring disordered domain formation on the stability of cholesterol-containing ordered domains (lipid rafts): identification of multiple raft-stabilization mechanisms. Biophys J 93: 4307–4318.

36. BrownDA, RoseJK (1992) Sorting of GPI-anchored proteins to glycolipid-enriched membrane subdomains during transport to the apical cell surface. Cell 68: 533–544.

37. LondonE, BrownDA (2000) Insolubility of lipids in triton X-100: physical origin and relationship to sphingolipid/cholesterol membrane domains (rafts). Biochim Biophys Acta 1508: 182–195.

38. PathakP, LondonE (2011) Measurement of lipid nanodomain (raft) formation and size in sphingomyelin/POPC/cholesterol vesicles shows TX-100 and transmembrane helices increase domain size by coalescing preexisting nanodomains but do not induce domain formation. Biophys J 101: 2417–2425.

39. BrownDA, LondonE (1998) Structure and origin of ordered lipid domains in biological membranes. J Membr Biol 164: 103–114.

40. WangTY, LeventisR, SilviusJR (2005) Artificially lipid-anchored proteins can elicit clustering-induced intracellular signaling events in Jurkat T-lymphocytes independent of lipid raft association. J Biol Chem 280: 22839–22846.

41. LaRoccaTJ, HolthausenDJ, HsiehC, RenkenC, MannellaCA, et al. (2009) The bactericidal effect of a complement-independent antibody is osmolytic and specific to Borrelia. Proc Natl Acad Sci U S A 106: 10752–10757.

42. Simon-PlasF, PerrakiA, BayerE, Gerbeau-PissotP, MongrandS (2011) An update on plant membrane rafts. Curr Opin Plant Biol 14: 642–649.

43. DupontS, BeneyL, FerreiraT, GervaisP (2011) Nature of sterols affects plasma membrane behavior and yeast survival during dehydration. Biochim Biophys Acta 1808: 1520–1528.

44. YeaglePL, MartinRB, LalaAK, LinHK, BlochK (1977) Differential effects of cholesterol and lanosterol on artificial membranes. Proc Natl Acad Sci U S A 74: 4924–4926.

45. ColemanJL, BenachJL (1992) Characterization of antigenic determinants of Borrelia burgdorferi shared by other bacteria. J Infect Dis 165: 658–666.

46. ColemanJL, KatonaLI, KuhlowC, ToledoA, OkanNA, et al. (2009) Evidence that two ATP-dependent (Lon) proteases in Borrelia burgdorferi serve different functions. PLoS Pathog 5: e1000676.

47. RipleyBD (1977) Modelling spatial patterns. Journal of the Royal Statistical Society Series B 39: 172–212.

48. RipleyBD (1979) Test of randomness for spatial point patterns. Journal of the Royal Statistical Society Series B 41: 368–374.

49. BesagJ, DigglePJ (1977) Simple Monte Carlo tests for spatial pattern. Applied Statistics 26: 327–333.

50. BlighEG, DyerWJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37: 911–917.

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Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

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