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  • 标题:A fast, single-vesicle fusion assay mimics physiological SNARE requirements
  • 本地全文:下载
  • 作者:Erdem Karatekin ; Jérôme Di Giovanni ; Cécile Iborra
  • 期刊名称:Proceedings of the National Academy of Sciences
  • 印刷版ISSN:0027-8424
  • 电子版ISSN:1091-6490
  • 出版年度:2010
  • 卷号:107
  • 期号:8
  • 页码:3517-3521
  • DOI:10.1073/pnas.0914723107
  • 语种:English
  • 出版社:The National Academy of Sciences of the United States of America
  • 摘要:Almost all known intracellular fusion reactions are driven by formation of trans-SNARE complexes through pairing of vesicle-associated v-SNAREs with complementary t-SNAREs on target membranes. However, the number of SNARE complexes required for fusion is unknown, and there is controversy about whether additional proteins are required to explain the fast fusion which can occur in cells. Here we show that single vesicles containing the synaptic/exocytic v-SNAREs VAMP/synaptobrevin fuse rapidly with planar, supported bilayers containing the synaptic/exocytic t-SNAREs syntaxin-SNAP25. Fusion rates decreased dramatically when the number of externally oriented v-SNAREs per vesicle was reduced below 5-10, directly establishing this as the minimum number required for rapid fusion. Docking-to-fusion delay time distributions were consistent with a requirement that 5-11 t-SNAREs be recruited to achieve fusion, closely matching the v-SNARE requirement.
  • 关键词:lipid bilayer ; membrane fusion ; SNARE mechanisms ; supported bilayer
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