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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Effects Of Linker Sequences On Vesicle Fusion Mediated By Lipid-anchored Dna Oligonucleotides
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Effects Of Linker Sequences On Vesicle Fusion Mediated By Lipid-anchored Dna Oligonucleotides

机译：接头序列对脂质锚定的Dna寡核苷酸介导的囊泡融合的影响。

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摘要

Synthetic lipid-oligonucleotide conjugates inserted into lipid vesicles mediate fusion when one population of vesicles displays the 5'-coupled conjugate and the other the 3-coupled conjugate, so that anti-parallel hybridization allows the membrane surfaces to come into close proximity. Improved assays show that lipid mixing proceeds more quickly and to a much greater extent than content mixing, suggesting the latter is rate limiting. To test the effect of membrane-membrane spacing on fusion, a series of conjugates was constructed by adding 2-24 noncomplementary bases at the membrane-proximal ends of two complementary sequences. Increasing linker lengths generally resulted in progressively reduced rates and extents of lipid and content mixing, in contrast to higher vesicle docking rates. The relatively flexible, single-stranded DNA linker facilitates docking but allows greater spacing between the vesicles after docking, thus making the transition into fusion less probable, but not preventing it altogether. These experiments demonstrate the utility of DNA as a model system for fusion proteins, where sequence can easily be modified to systematically probe the effect of distance between bilayers in the fusion reaction.
机译：当一个囊泡群显示5'-偶联的偶联物，而另一个囊泡3-偶联的偶联物时，插入脂质囊泡的合成脂质-寡核苷酸偶联物可介导融合，因此反平行杂交可使膜表面紧密接近。改进的试验表明，脂质混合比内容混合更快，更大程度地进行，这表明后者是速率限制的。为了测试膜-膜间隔对融合的影响，通过在两个互补序列的膜近端添加2-24个非互补碱基来构建一系列结合物。与较高的囊泡对接速率相反，增加的接头长度通常导致脂质和内含物混合的速率和程度逐渐降低。相对灵活的单链DNA接头有利于对接，但对接后囊泡之间的间隔更大，从而使向融合的转变不太可能，但不能完全阻止融合。这些实验证明了DNA作为融合蛋白的模型系统的实用性，在该系统中可以轻松地修饰序列以系统地探测融合反应中双层之间距离的影响。

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