#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

Botulinum Neurotoxin D Uses Synaptic Vesicle Protein SV2 and Gangliosides as Receptors


Botulinum neurotoxins (BoNTs) include seven bacterial toxins (BoNT/A-G) that target presynaptic terminals and act as proteases cleaving proteins required for synaptic vesicle exocytosis. Here we identified synaptic vesicle protein SV2 as the protein receptor for BoNT/D. BoNT/D enters cultured hippocampal neurons via synaptic vesicle recycling and can bind SV2 in brain detergent extracts. BoNT/D failed to bind and enter neurons lacking SV2, which can be rescued by expressing one of the three SV2 isoforms (SV2A/B/C). Localization of SV2 on plasma membranes mediated BoNT/D binding in both neurons and HEK293 cells. Furthermore, chimeric receptors containing the binding sites for BoNT/A and E, two other BoNTs that use SV2 as receptors, failed to mediate the entry of BoNT/D suggesting that BoNT/D binds SV2 via a mechanism distinct from BoNT/A and E. Finally, we demonstrated that gangliosides are essential for the binding and entry of BoNT/D into neurons and for its toxicity in vivo, supporting a double-receptor model for this toxin.


Vyšlo v časopise: Botulinum Neurotoxin D Uses Synaptic Vesicle Protein SV2 and Gangliosides as Receptors. PLoS Pathog 7(3): e32767. doi:10.1371/journal.ppat.1002008
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1002008

Souhrn

Botulinum neurotoxins (BoNTs) include seven bacterial toxins (BoNT/A-G) that target presynaptic terminals and act as proteases cleaving proteins required for synaptic vesicle exocytosis. Here we identified synaptic vesicle protein SV2 as the protein receptor for BoNT/D. BoNT/D enters cultured hippocampal neurons via synaptic vesicle recycling and can bind SV2 in brain detergent extracts. BoNT/D failed to bind and enter neurons lacking SV2, which can be rescued by expressing one of the three SV2 isoforms (SV2A/B/C). Localization of SV2 on plasma membranes mediated BoNT/D binding in both neurons and HEK293 cells. Furthermore, chimeric receptors containing the binding sites for BoNT/A and E, two other BoNTs that use SV2 as receptors, failed to mediate the entry of BoNT/D suggesting that BoNT/D binds SV2 via a mechanism distinct from BoNT/A and E. Finally, we demonstrated that gangliosides are essential for the binding and entry of BoNT/D into neurons and for its toxicity in vivo, supporting a double-receptor model for this toxin.


Zdroje

1. SchiavoGMatteoliMMontecuccoC 2000 Neurotoxins affecting neuroexocytosis. Physiol Rev 80 717 766

2. ArnonSSSchechterRInglesbyTVHendersonDABartlettJG 2001 Botulinum toxin as a biological weapon: medical and public health management. JAMA 285 1059 1070

3. SimpsonLL 1986 Molecular pharmacology of botulinum toxin and tetanus toxin. Annu Rev Pharmacol Toxicol 26 427 453

4. MontalM 2010 Botulinum neurotoxin: a marvel of protein design. Annu Rev Biochem 79 591 617

5. SchantzEJJohnsonEA 1992 Properties and use of botulinum toxin and other microbial neurotoxins in medicine. Microbiol Rev 56 80 99

6. JohnsonEA 1999 Clostridial toxins as therapeutic agents: benefits of nature's most toxic proteins. Annu Rev Microbiol 53 551 575

7. MontecuccoCMolgoJ 2005 Botulinal neurotoxins: revival of an old killer. Curr Opin Pharmacol 5 274 279

8. DollyJOLawrenceGWMengJWangJ 2009 Neuro-exocytosis: botulinum toxins as inhibitory probes and versatile therapeutics. Curr Opin Pharmacol 9 326 335

9. HillKKSmithTJHelmaCHTicknorLOFoleyBT 2007 Genetic diversity among Botulinum Neurotoxin-producing clostridial strains. J Bacteriol 189 818 832

10. SimpsonLL 2004 Identification of the major steps in botulinum toxin action. Annu Rev Pharmacol Toxicol 44 167 193

11. SchiavoGBenfenatiFPoulainBRossettoOPolverino de LauretoP 1992 Tetanus and botulinum-B neurotoxins block neurotransmitter release by proteolytic cleavage of synaptobrevin. Nature 359 832 835

12. SchiavoGRossettoOCatsicasSPolverino de LauretoPDasGuptaBR 1993 Identification of the nerve terminal targets of botulinum neurotoxin serotypes A, D, and E. J Biol Chem 268 23784 23787

13. SchiavoGShoneCCRossettoOAlexanderFCMontecuccoC 1993 Botulinum neurotoxin serotype F is a zinc endopeptidase specific for VAMP/synaptobrevin. J Biol Chem 268 11516 11519

14. SchiavoGMalizioCTrimbleWSPolverino de LauretoPMilanG 1994 Botulinum G neurotoxin cleaves VAMP/synaptobrevin at a single Ala-Ala peptide bond. J Biol Chem 269 20213 20216

15. YamasakiSBinzTHayashiTSzaboEYamasakiN 1994 Botulinum neurotoxin type G proteolyses the Ala81-Ala82 bond of rat synaptobrevin 2. Biochem Biophys Res Commun 200 829 835

16. YamasakiSBaumeisterABinzTBlasiJLinkE 1994 Cleavage of members of the synaptobrevin/VAMP family by types D and F botulinal neurotoxins and tetanus toxin. J Biol Chem 269 12764 12772

17. BlasiJChapmanERLinkEBinzTYamasakiS 1993 Botulinum neurotoxin A selectively cleaves the synaptic protein SNAP-25. Nature 365 160 163

18. SchiavoGSantucciADasguptaBRMehtaPPJontesJ 1993 Botulinum neurotoxins serotypes A and E cleave SNAP-25 at distinct COOH-terminal peptide bonds. FEBS Lett 335 99 103

19. WilliamsonLCHalpernJLMontecuccoCBrownJENealeEA 1996 Clostridial neurotoxins and substrate proteolysis in intact neurons: botulinum neurotoxin C acts on synaptosomal-associated protein of 25 kDa. J Biol Chem 271 7694 7699

20. ForanPLawrenceGWShoneCCFosterKADollyJO 1996 Botulinum neurotoxin C1 cleaves both syntaxin and SNAP-25 in intact and permeabilized chromaffin cells: correlation with its blockade of catecholamine release. Biochemistry 35 2630 2636

21. BinzTBlasiJYamasakiSBaumeisterALinkE 1994 Proteolysis of SNAP-25 by types E and A botulinal neurotoxins. J Biol Chem 269 1617 1620

22. SchiavoGShoneCCBennettMKSchellerRHMontecuccoC 1995 Botulinum neurotoxin type C cleaves a single Lys-Ala bond within the carboxyl-terminal region of syntaxins. J Biol Chem 270 10566 10570

23. BlasiJChapmanERYamasakiSBinzTNiemannH 1993 Botulinum neurotoxin C1 blocks neurotransmitter release by means of cleaving HPC-1/syntaxin. EMBO J 12 4821 4828

24. SollnerTWhiteheartSWBrunnerMErdjument-BromageHGeromanosS 1993 SNAP receptors implicated in vesicle targeting and fusion. Nature 362 318 324

25. WeberTZemelmanBVMcNewJAWestermannBGmachlM 1998 SNAREpins: minimal machinery for membrane fusion. Cell 92 759 772

26. JahnRSchellerRH 2006 SNAREs—engines for membrane fusion. Nat Rev Mol Cell Biol 7 631 643

27. SuttonRBFasshauerDJahnRBrungerAT 1998 Crystal structure of a SNARE complex involved in synaptic exocytosis at 2.4 A resolution. Nature 395 347 353

28. SudhofTCRothmanJE 2009 Membrane fusion: grappling with SNARE and SM proteins. Science 323 474 477

29. MontecuccoCRossettoOSchiavoG 2004 Presynaptic receptor arrays for clostridial neurotoxins. Trends Microbiol 12 442 446

30. BinzTRummelA 2009 Cell entry strategy of clostridial neurotoxins. J Neurochem 109 1584 1595

31. SimpsonLLRapportMM 1971 Ganglioside inactivation of botulinum toxin. J Neurochem 18 1341 1343

32. KitamuraMIwamoriMNagaiY 1980 Interaction between Clostridium botulinum neurotoxin and gangliosides. Biochim Biophys Acta 628 328 335

33. KamataYKozakiSSakaguchiGIwamoriMNagaiY 1986 Evidence for direct binding of Clostridium botulinum type E derivative toxin and its fragments to gangliosides and free fatty acids. Biochem Biophys Res Commun 140 1015 1019

34. KozakiSKamataYWataraiSNishikiTMochidaS 1998 Ganglioside GT1b as a complementary receptor component for Clostridium botulinum neurotoxins. Microb Pathog 25 91 99

35. RummelAMahrholdSBigalkeHBinzT 2004 The HCC-domain of botulinum neurotoxins A and B exhibits a singular ganglioside binding site displaying serotype specific carbohydrate interaction. Mol Microbiol 51 631 643

36. YowlerBCSchengrundCL 2004 Botulinum neurotoxin A changes conformation upon binding to ganglioside GT1b. Biochemistry 43 9725 9731

37. SwaminathanSEswaramoorthyS 2000 Structural analysis of the catalytic and binding sites of Clostridium botulinum neurotoxin B. Nat Struct Biol 7 693 699

38. KohdaTIharaHSetoYTsutsukiHMukamotoM 2007 Differential contribution of the residues in C-terminal half of the heavy chain of botulinum neurotoxin type B to its binding to the ganglioside GT1b and the synaptotagmin 2/GT1b complex. Microb Pathog 42 72 79

39. StenmarkPDupuyJImamuraAKisoMStevensRC 2008 Crystal structure of botulinum neurotoxin type A in complex with the cell surface co-receptor GT1b-insight into the toxin-neuron interaction. PLoS Pathog 4 e1000129

40. StenmarkPDongMDupuyJChapmanERStevensRC 2010 Crystal structure of the botulinum neurotoxin type G binding domain: insight into cell surface binding. J Mol Biol 397 1287 1297

41. SchmittJKaralewitzABenefieldDAMushrushDJPruittRN 2010 Structural analysis of botulinum neurotoxin type G receptor binding. Biochemistry 49 5200 5205

42. StrotmeierJLeeKVolkerAKMahrholdSZongY 2010 Botulinum neurotoxin serotype D attacks neurons via two carbohydrate binding sites in a ganglioside dependent manner. Biochem J 431 207 216

43. RummelAHafnerKMahrholdSDarashchonakNHoltM 2009 Botulinum neurotoxins C, E and F bind gangliosides via a conserved binding site prior to stimulation-dependent uptake with botulinum neurotoxin F utilising the three isoforms of SV2 as second receptor. J Neurochem 110 1942 1954

44. KaralewitzAPKrokenARFuZBaldwinMRKimJJ 2010 Identification of a Unique Ganglioside Binding Loop within Botulinum Neurotoxins C and D-SA. Biochemistry 49 8117 8126

45. FuZChenCBarbieriJTKimJJBaldwinMR 2009 Glycosylated SV2 and gangliosides as dual receptors for botulinum neurotoxin serotype F. Biochemistry 48 5631 5641

46. RummelAEichnerTWeilTKarnathTGutcaitsA 2007 Identification of the protein receptor binding site of botulinum neurotoxins B and G proves the double-receptor concept. Proc Natl Acad Sci U S A 104 359 364

47. YowlerBCKensingerRDSchengrundCL 2002 Botulinum neurotoxin A activity is dependent upon the presence of specific gangliosides in neuroblastoma cells expressing synaptotagmin I. J Biol Chem 277 32815 32819

48. ChaiQArndtJWDongMTeppWHJohnsonEA 2006 Structural basis of cell surface receptor recognition by botulinum neurotoxin B. Nature 444 1096 1100

49. DongMTeppWHLiuHJohnsonEAChapmanER 2007 Mechanism of botulinum neurotoxin B and G entry into hippocampal neurons. J Cell Biol 179 1511 1522

50. BullensRWO'HanlonGMWagnerEMolenaarPCFurukawaK 2002 Complex gangliosides at the neuromuscular junction are membrane receptors for autoantibodies and botulinum neurotoxin but redundant for normal synaptic function. J Neurosci 22 6876 6884

51. DongMLiuHTeppWHJohnsonEAJanzR 2008 Glycosylated SV2A and SV2B mediate the entry of botulinum neurotoxin E into neurons. Mol Biol Cell 19 5226 5237

52. KitamuraMTakamiyaKAizawaSFurukawaK 1999 Gangliosides are the binding substances in neural cells for tetanus and botulinum toxins in mice. Biochim Biophys Acta 1441 1 3

53. TsukamotoKKohdaTMukamotoMTakeuchiKIharaH 2005 Binding of Clostridium botulinum type C and D neurotoxins to ganglioside and phospholipid. Novel insights into the receptor for clostridial neurotoxins. J Biol Chem 280 35164 35171

54. MontecuccoC 1986 How do tetanus and botulinum toxins bind to neuronal membranes? Trends Biochem Sci 314 317

55. NishikiTKamataYNemotoYOmoriAItoT 1994 Identification of protein receptor for Clostridium botulinum type B neurotoxin in rat brain synaptosomes. J Biol Chem 269 10498 10503

56. NishikiTTokuyamaYKamataYNemotoYYoshidaA 1996 The high-affinity binding of Clostridium botulinum type B neurotoxin to synaptotagmin II associated with gangliosides GT1b/GD1a. FEBS Lett 378 253 257

57. DongMRichardsDAGoodnoughMCTeppWHJohnsonEA 2003 Synaptotagmins I and II mediate entry of botulinum neurotoxin B into cells. J Cell Biol 162 1293 1303

58. RummelAKarnathTHenkeTBigalkeHBinzT 2004 Synaptotagmins I and II act as nerve cell receptors for botulinum neurotoxin G. J Biol Chem 279 30865 30870

59. JinRRummelABinzTBrungerAT 2006 Botulinum neurotoxin B recognizes its protein receptor with high affinity and specificity. Nature 444 1092 1095

60. DongMYehFTeppWHDeanCJohnsonEA 2006 SV2 is the protein receptor for botulinum neurotoxin A. Science 312 592 596

61. MahrholdSRummelABigalkeHDavletovBBinzT 2006 The synaptic vesicle protein 2C mediates the uptake of botulinum neurotoxin A into phrenic nerves. FEBS Lett 580 2011 2014

62. BajjaliehSMPetersonKShinghalRSchellerRH 1992 SV2, a brain synaptic vesicle protein homologous to bacterial transporters. Science 257 1271 1273

63. FeanyMBLeeSEdwardsRHBuckleyKM 1992 The synaptic vesicle protein SV2 is a novel type of transmembrane transporter. Cell 70 861 867

64. BajjaliehSMPetersonKLinialMSchellerRH 1993 Brain contains two forms of synaptic vesicle protein 2. Proc Natl Acad Sci U S A 90 2150 2154

65. JanzRSudhofTC 1999 SV2C is a synaptic vesicle protein with an unusually restricted localization: anatomy of a synaptic vesicle protein family. Neuroscience 94 1279 1290

66. LacyDBStevensRC 1999 Sequence homology and structural analysis of the clostridial neurotoxins. J Mol Biol 291 1091 1104

67. DollyJOBlackJWilliamsRSMellingJ 1984 Acceptors for botulinum neurotoxin reside on motor nerve terminals and mediate its internalization. Nature 307 457 460

68. SudhofTC 2004 The synaptic vesicle cycle. Annu Rev Neurosci 27 509 547

69. TakamoriSHoltMSteniusKLemkeEAGronborgM 2006 Molecular anatomy of a trafficking organelle. Cell 127 831 846

70. BennettMKCalakosNKreinerTSchellerRH 1992 Synaptic vesicle membrane proteins interact to form a multimeric complex. J Cell Biol 116 761 775

71. SchivellAEBatchelorRHBajjaliehSM 1996 Isoform-specific, calcium-regulated interaction of the synaptic vesicle proteins SV2 and synaptotagmin. J Biol Chem 271 27770 27775

72. LazzellDRBelizaireRThakurPSherryDMJanzR 2004 SV2B regulates synaptotagmin 1 by direct interaction. J Biol Chem 279 52124 52131

73. BaldwinMRBarbieriJT 2007 Association of botulinum neurotoxin serotypes a and B with synaptic vesicle protein complexes. Biochemistry 46 3200 3210

74. YaoJNowackAKensel-HammesPGardnerRGBajjaliehSM 2010 Cotrafficking of SV2 and synaptotagmin at the synapse. J Neurosci 30 5569 5578

75. YamashitaTHashiramotoAHaluzikMMizukamiHBeckS 2003 Enhanced insulin sensitivity in mice lacking ganglioside GM3. Proc Natl Acad Sci U S A 100 3445 3449

76. LiuYWadaRKawaiHSangoKDengC 1999 A genetic model of substrate deprivation therapy for a glycosphingolipid storage disorder. J Clin Invest 103 497 505

77. BoroffDAFleckU 1966 Statistical analysis of a rapid in vivo method for the titration of the toxin of Clostridium botulinum. J Bacteriol 92 1580 1581

78. LacyDBTeppWCohenACDasGuptaBRStevensRC 1998 Crystal structure of botulinum neurotoxin type A and implications for toxicity. Nat Struct Biol 5 898 902

79. KumaranDEswaramoorthySFureyWNavazaJSaxM 2009 Domain organization in Clostridium botulinum neurotoxin type E is unique: its implication in faster translocation. J Mol Biol 386 233 245

80. ChapmanER 2002 Synaptotagmin: a Ca(2+) sensor that triggers exocytosis? Nat Rev Mol Cell Biol 3 498 508

81. ZhangJZDavletovBASudhofTCAndersonRG 1994 Synaptotagmin I is a high affinity receptor for clathrin AP-2: implications for membrane recycling. Cell 78 751 760

82. HauckeVDe CamilliP 1999 AP-2 recruitment to synaptotagmin stimulated by tyrosine-based endocytic motifs. Science 285 1268 1271

83. Nicholson-TomishimaKRyanTA 2004 Kinetic efficiency of endocytosis at mammalian CNS synapses requires synaptotagmin I. Proc Natl Acad Sci U S A 101 16648 16652

84. PoskanzerKEMarekKWSweeneySTDavisGW 2003 Synaptotagmin I is necessary for compensatory synaptic vesicle endocytosis in vivo. Nature 426 559 563

85. WanQFZhouZYThakurPVilaASherryDM 2010 SV2 acts via presynaptic calcium to regulate neurotransmitter release. Neuron 66 884 895

86. DirilMKWienischMJungNKlingaufJHauckeV 2006 Stonin 2 is an AP-2-dependent endocytic sorting adaptor for synaptotagmin internalization and recycling. Dev Cell 10 233 244

87. EleopraRTugnoliVRossettoOMontecuccoCDe GrandisD 1997 Botulinum neurotoxin serotype C: a novel effective botulinum toxin therapy in human. Neurosci Lett 224 91 94

88. EleopraRTugnoliVQuatraleRRossettoOMontecuccoC 2006 Clinical use of non-A botulinum toxins: botulinum toxin type C and botulinum toxin type F. Neurotox Res 9 127 131

89. WebbRPSmithTJWrightPMMontgomeryVAMeagherMM 2007 Protection with recombinant Clostridium botulinum C1 and D binding domain subunit (Hc) vaccines against C and D neurotoxins. Vaccine 25 4273 4282

90. JanzRGodaYGeppertMMisslerMSudhofTC 1999 SV2A and SV2B function as redundant Ca2+ regulators in neurotransmitter release. Neuron 24 1003 1016

91. JanzRSudhofTCHammerREUnniVSiegelbaumSA 1999 Essential roles in synaptic plasticity for synaptogyrin I and synaptophysin I. Neuron 24 687 700

92. GasconSPaez-GomezJADiaz-GuerraMScheiffelePSchollFG 2008 Dual-promoter lentiviral vectors for constitutive and regulated gene expression in neurons. J Neurosci Methods 168 104 112

93. ChapmanERJahnR 1994 Calcium-dependent interaction of the cytoplasmic region of synaptotagmin with membranes. Autonomous function of a single C2-homologous domain. J Biol Chem 269 5735 5741

94. ChapmanERAnSEdwardsonJMJahnR 1996 A novel function for the second C2 domain of synaptotagmin. Ca2+-triggered dimerization. J Biol Chem 271 5844 5849

95. LewisJLDongMEarlesCAChapmanER 2001 The transmembrane domain of syntaxin 1A is critical for cytoplasmic domain protein-protein interactions. J Biol Chem 276 15458 15465

Štítky
Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

Článok vyšiel v časopise

PLOS Pathogens


2011 Číslo 3
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Aktuální možnosti diagnostiky a léčby litiáz
nový kurz
Autori: MUDr. Tomáš Ürge, PhD.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa

#ADS_BOTTOM_SCRIPTS#