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首页> 外文期刊>Biophysical Chemistry: An International Journal Devoted to the Physical Chemistry of Biological Phenomena >Membrane protein reconstitution into liposomes guided by dual-color fluorescence cross-correlation spectroscopy
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Membrane protein reconstitution into liposomes guided by dual-color fluorescence cross-correlation spectroscopy

机译：膜蛋白重构成脂质体以双色荧光互相关光谱引导

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摘要

Proteoliposomes represent nanoscale assemblies of indispensable value for studying membrane proteins in general and membrane transporters in particular. Since no universal protocol exists, conditions for proteoliposome formation must be determined on a case-by-case basis. This process will be significantly expedited if the size and composition of the assemblies can be analyzed in a single step using only microliters of sample. Here we show that dual-color fluorescence cross-correlation spectroscopy (FCCS) is of great value for optimizing the re-constitution process, because it distinguishes micelles, liposomes and aggregates in heterogeneous mixtures and permits direct monitoring of the co-localization of proteins and lipids in the diffusing assemblies. As proof-of-principle, liposomes containing the functional multidrug resistance transporter NorA from Staphylococcus au-reus were prepared, demonstrating that FCCS is an excellent tool to guide the development of reconstitution protocols.
机译：蛋白环体代表纳米级组件，用于研究一般和膜转运蛋白的膜蛋白的不可或缺的价值。由于不存在通用方案，因此必须根据案例确定蛋白质体形成的条件。如果组件的尺寸和组成可以在仅使用微升样品中可以在单一的步骤中分析组件的尺寸和组成，将显着加速该方法。在这里，我们表明双色荧光互相关光谱（FCCS）具有很大的优化重新构造过程的价值，因为它将胶束，脂质体和聚集体区分开在异质混合物中，并且允许直接监测蛋白质的共定位和蛋白质的共定位弥漫性组件中的脂质。作为原则上，制备了含有功能性多药耐药耐药术Nora的原则上，从而制备了来自葡萄球菌的葡萄球菌，表明FCCS是引导重构方案发展的优异工具。

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