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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Kinetics of DNA-mediated docking reactions between vesicles tethered to supported lipid bilayers
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Kinetics of DNA-mediated docking reactions between vesicles tethered to supported lipid bilayers

机译：连接到支持的脂质双层的囊泡之间DNA介导的对接反应的动力学

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摘要

Membrane-membrane recognition and binding are crucial in many biological processes. We report an approach to studying the dynamics of such reactions by using DNA-tethered vesicles as a general scaffold for displaying membrane components. This system was used to characterize the docking reaction between two populations of tethered vesicles that display complementary DNA. Deposition of vesicles onto a supported lipid bilayer was performed by using a microfluidic device to prevent mixing of the vesicles in bulk during sample preparation. Once tethered onto the surface, vesicles mixed via two-dimensional diffusion. DNA-mediated docking of two reacting vesicles results in their colocalization after collision and their subsequent tandem motion. Individual docking events and population kinetics were observed via epiflu-orescence microscopy. A lattice-diffusion simulation was implemented to extract from experimental data the probability, Pdock, that a collision leads to docking. For individual vesicles displaying small numbers of docking DNA, Pdock shows a first-order relationship with copy number as well as a strong dependence on the DNA sequence. Both trends are explained by a model that includes both tethered vesicle diffusion on the supported bilayer and docking DNA diffusion over each vesicle's surface. These results provide the basis for the application of tethered vesicles to study other membrane reactions including protein-mediated docking and fusion.
机译：膜-膜的识别和结合在许多生物学过程中至关重要。我们报告了一种方法，通过使用DNA束缚的囊泡作为展示膜成分的通用支架，来研究此类反应的动力学。该系统用于表征显示互补DNA的两个束缚水泡群体之间的对接反应。通过使用微流体装置将囊泡沉积到支持的脂质双层上，以防止在样品制备过程中大量囊泡混合。一旦束缚在表面上，囊泡通过二维扩散混合。 DNA介导的两个反应小泡的对接导致它们在碰撞后共定位并随后进行串联运动。通过流行荧光显微镜观察到个体对接事件和种群动力学。实施了晶格扩散模拟，以从实验数据中提取碰撞导致对接的概率Pdock。对于显示少量对接DNA的单个囊泡，Pdock显示与拷贝数的一阶关系以及对DNA序列的强烈依赖性。两种趋势都可以通过一个模型来解释，该模型既包括在支持的双层分子上的束缚小泡扩散，又包括每个小泡表面上的DNA扩散对接。这些结果为拴系小泡用于研究其他膜反应包括蛋白介导的对接和融合提供了基础。

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