Rapid SNARE-Mediated Fusion of Liposomes and Chromaffin Granules with Giant Unilamellar Vesicles.

Rapid SNARE-Mediated Fusion of Liposomes and Chromaffin Granules with Giant Unilamellar Vesicles.
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DOI:
10.1016/j.bpj.2017.03.010
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发表时间:
2017-09-19
影响因子:
3.4
通讯作者:
Jahn R
Jahn R
中科院分区:
生物学3区
文献类型:
--
作者:
Witkowska A;Jahn R

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可溶性N-乙基马来酰亚胺敏感因子激活蛋白受体(SNARE)蛋白是真核细胞分泌途径中膜融合的主要催化剂。在体外,SNARE 足以介导天然膜和人造膜的有效融合。据我们所知,我们在这里建立了一个新的平台,用于监测巨型单层囊泡(GUV)与较小的脂质体或纯化的分泌颗粒之间的对接和融合,具有高时间和空间分辨率。融合分析仅限于 GUV 膜的独立部分,该部分表现出低曲率且缺乏表面接触,从而避免了与支持的双层或表面固定的小囊泡融合时粘附介导的对融合反应的干扰。我们的结果表明,脂质体和嗜铬颗粒与含有激活的 SNARE 的 GUV 融合,对接和融合之间只有几毫秒的延迟。我们得出的结论是,在反式初次接触后,单独的 SNARE 就可以以接近快速神经元胞吐作用的速度完成融合。
Soluble N-ethylmaleimide-sensitive factor activating protein receptor (SNARE) proteins are the main catalysts for membrane fusion in the secretory pathway of eukaryotic cells. In vitro, SNAREs are sufficient to mediate effective fusion of both native and artificial membranes. Here we have established, to our knowledge, a new platform for monitoring SNARE-mediated docking and fusion between giant unilamellar vesicles (GUVs) and smaller liposomes or purified secretory granules with high temporal and spatial resolution. Analysis of fusion is restricted to the free-standing part of the GUV-membrane exhibiting low curvature and a lack of surface contact, thus avoiding adhesion-mediated interference with the fusion reaction as in fusion with supported bilayers or surface-immobilized small vesicles. Our results show that liposomes and chromaffin granules fuse with GUVs containing activated SNAREs with only few milliseconds delay between docking and fusion. We conclude that after initial contact in trans, SNAREs alone can complete fusion at a rate close to fast neuronal exocytosis.
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