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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Two synaptobrevin molecules are sufficient for vesicle fusion in central nervous system synapses
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Two synaptobrevin molecules are sufficient for vesicle fusion in central nervous system synapses

机译：两个突触短纤维蛋白分子足以在中枢神经系统突触中进行囊泡融合

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摘要

Exocytosis of synaptic vesicles (SVs) during fast synaptic transmission is mediated by soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex assembly formed by the coil-coiling of three members of this protein family: vesicle SNARE protein, synaptobrevin 2 (syb2), and the presynaptic membrane SNAREs syntaxin-1A and SNAP-25. However, it is controversially debated how many SNARE complexes are minimally needed for SV priming and fusion. To quantify this effective number, we measured the fluorescence responses from single fusing vesicles expressing pHluorin (pHI), a pH-sensitive variant of GFP, fused to the luminal domain of the vesicular SNARE syb2 (spH) in cultured hippocampal neurons lacking endogenous syb2. Fluorescence responses were quantal, with the unitary signals precisely corresponding to single pHluorin molecules. Using this approach we found that two copies of spH per SV fully rescued evoked fusion whereas SVs expressing only one spH were unable to rapidly fuse upon stimulation. Thus, two syb2 molecules and likely two SNARE complexes are necessary and sufficient for SV fusion during fast synaptic transmission.
机译：突触小泡（SVs）在突触快速传递过程中的胞吐作用是由可溶性N-乙基马来酰亚胺敏感因子附着蛋白受体（SNARE）复杂复合体介导的，该复合体由该蛋白家族的三个成员的卷曲螺旋形成：囊泡SNARE蛋白，突触连接蛋白2（ syb2）和突触前膜SNAREs语法in-1A和SNAP-25。然而，有争议的是，SV引发和融合最低需要多少个SNARE复合物。为了量化此有效数目，我们测量了表达pHluorin（pHI）的单个融合囊泡的荧光响应，pHluorin是GFP的pH敏感变体，与缺乏内源性syb2的海马神经元的囊泡SNARE syb2（spH）的腔结构域融合。荧光响应是定量的，单位信号精确对应于单个pHluorin分子。使用这种方法，我们发现每个SV有两个拷贝的spH完全拯救了诱发的融合，而仅表达一个spH的SV在刺激后无法快速融合。因此，在快速突触传递过程中，两个syb2分子以及可能的两个SNARE复合物对于SV融合是必需的和充分的。

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  • 作者

    Raunak Sinha; Saheeb Ahmed; Reinhard Jahn; Jurgen Klingauf;

  • 作者单位

    Departments of Membrane Biophysics, Max-Planck Institute for Biophysical Chemistry, 37077 Gottingen, Germany,Department of Cellular Biophysics, Institute for Medical Physics and Biophysics, University of Muenster, 48149 Muenster, Germany;

    Departments of Neurobiology, Max-Planck Institute for Biophysical Chemistry, 37077 Gottingen, Germany;

    Departments of Neurobiology, Max-Planck Institute for Biophysical Chemistry, 37077 Gottingen, Germany;

    Departments of Membrane Biophysics, Max-Planck Institute for Biophysical Chemistry, 37077 Gottingen, Germany,Department of Cellular Biophysics, Institute for Medical Physics and Biophysics, University of Muenster, 48149 Muenster, Germany;

  • 收录信息 美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    synaptophluorin; membrane fusion; single molecule bleaching; SNARE density;

    机译：突触核蛋白膜融合单分子漂白网罗密度;

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