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Imaging Fast SNARE Mediated-Membrane Fusion in Planar-Supported Bilayers Most conspicuous are differences in composition of the target membranes.Fix et al. (6) used synthetic POPC lipids and a higher density (lipid/protein:;3000) of coexpressed syntaxin1A/SNAP25 complex. Fusion was triggered by adding Ca21 ions in their assay. Bowen et al. (7) used extracted lipid mixtures of eggPC and brainPS and a low density (lipid/protein: ;14,000) of syntaxin in their membranes and triggered fusion thermally. Liu et al. were able to achieve high fusion rates without an external trigger when they used a very low density (lipid/protein:;30,000) of either syntaxin1A or syntaxin1A/SNAP25 complexes in a membrane consisting of the synthetic lipids POPC and DOPS. Although the authors showed that a high density of t-SNAREs prevents fusion in their system, the influence of the lipid environment was not studied systematically. By comparing the experiments of the different groups, it seems that the lipid composition indeed plays a crucial role in vesicle membrane fusion. The observation of fast fusion events coupled with rates of lipid diffusion similar to those observed in cell membranes are evidence for the high quality of Liu et al.’s planar-supported membranes and prove that this constitutes an excellent experimental system to address questions that remain to more fully understand exocytotic membrane fusion at the molecular mechanistic level. The variation of lipid and protein compositions as well as the use of membrane and content labels should further unveil details about the molecular mechanisms and kinetics of pore formation during membrane fusion. It will be interesting to see if further perfection of this assay will resolve remaining questions about the role of SNAP25 and other accessory proteins in synaptic membrane fusion and eventually raise the fusion rate further to those observed in cells. [ Last edited by hzxhmomo on 2008-5-15 at 09:15 ] |
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flyrain363
木虫 (正式写手)
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有些句子好像拗口，不知道是否符合要求
★ ★ ★ ★ ★ ★ ★ ★ ★ ★
hzxhmomo(金币+10,VIP+0):是有些拗的,所以翻啊翻不出,谢谢
hzxhmomo(金币+10,VIP+0):是有些拗的,所以翻啊翻不出,谢谢
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基于平面双层膜分析的SNARE快速介导膜融合的成像研究 最显著的区别在于目标膜组分的差异。Fix 等人使用合成的POPC脂质和共表达syntaxinA/SNAP25混合物的高密度脂蛋白（脂质/蛋白：；3000）进行研究。在他们的研究中，加入Ca21 离子则促发融合的发生。Bowen 等人使用鸡蛋PC和脑PS提取的脂质混合物与含syntaxin的低密度脂蛋白（脂质/蛋白：；14，000）研究发现，加热可以促发膜融合。Liu等人在使用非常低密度（脂质/蛋白：；30，000）的混合物时，无论syntaxin1A还是syntaxin1A/SNAP25混合物中含有合成的脂质POPC和DOPS时，无需额外促发均可以实现高融合率。虽然作者表明，在他们的研究系统中，高密度的t-SNAREs可以防止融合的发生，但是脂质所处环境对融合的影响却没有系统的研究。比较不同的研究结果，似乎不同的脂质组分对囊泡膜融合具有至关重要的作用。能够观察到具有活细胞中脂质扩散速率的快速融合，是Liu等人基于平面双层膜分析实验的高可靠性的证据。这就构成了一个很好的实验系统，便于在分子水平更好地了解胞吐过程中遗留的问题。 脂质和蛋白组分的变化以及膜和示踪标签（组分标签？）的使用，可以更加详细的了解在膜融合过程中的分子机制以及孔隙形成时的动力学变化。如果进一步完善，该方法将有助于了解在突触膜融合中snap25和其他配件蛋白发挥的其他未知作用，并最终获得比活细胞中更高的融合效率。 |
2楼2008-05-15 23:17:25
