botulinum neurotoxin d uses synaptic vesicle protein sv2 and gangliosides as receptors肉毒神经毒素d使用突触囊泡蛋白sv2和神经节甘脂作为受体.pdfVIP

Botulinum Neurotoxin D Uses Synaptic Vesicle Protein SV2 and Gangliosides as Receptors 1 2 2 1 Lisheng Peng , William H. Tepp , Eric A. Johnson , Min Dong * 1 Department of Microbiology and Molecular Genetics, Harvard Medical School and Division of Neuroscience, New England Primate Research Center, Southborough, Massachusetts, United States of America, 2 Department of Food Microbiology and Toxicology, University of Wisconsin, Madison, Wisconsin, United States of America Abstract 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. Citation: Peng L, Tepp WH, Johnson EA, Dong M (2011) Botulinum Neurotoxin D Uses Synaptic Vesicle Protein SV2 and Gangliosides as Receptors. PLoS Pathog 7(3): e1002008. doi:10.1371/journal.ppat.1002008 Editor: Theresa Koehler, The University of Texas-Hou