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Introducing SEC-SANS for studies of complex self-organized biological systems

机译:引入SEC-SANS复杂的研究自组织的生物系统

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Small-angle neutron scattering (SANS) is maturing as a method for studying complex biological structures. Owing to the intrinsic ability of the technique to discern between ~1H- and ~2H-labelled particles, it is especially useful for contrast-variation studies of biological systems containing multiple components. SANS is complementary to small-angle X-ray scattering (SAXS), in which similar contrast variation is not easily performed but in which data with superior counting statistics are more easily obtained. Obtaining small-angle scattering (SAS) data on monodisperse complex biological structures is often challenging owing to sample degradation and/or aggregation. This problem is enhanced in the D_2O-based buffers that are typically used in SANS. In SAXS, such problems are solved using an online size-exclusion chromatography (SEC) setup. In the present work, the feasibility of SEC-SANS was investigated using a series of complex and difficult samples of membrane proteins embedded in nanodisc particles that consist of both phospholipid and protein components. It is demonstrated that SEC-SANS provides data of sufficient signal-to-noise ratio for these systems, while at the same time circumventing aggregation. By combining SEC-SANS and SEC-SAXS data, an optimized basis for refining structural models of the investigated structures is obtained.
机译:小角中子散射(SANS)是成熟作为一个方法来研究复杂的生物结构。技术来辨别和~ 1 h -之间~ 2 h-labelled粒子,它尤其有用contrast-variation研究的生物系统包含多个组件。小角x射线散射的补充(粉煤灰),类似的对比变化不容易但在执行数据优越的计算统计数据更容易获得的。单分散的数据复杂的生物由于样本结构通常是具有挑战性的退化和/或聚合。增强D_2O-based缓冲区中通常用于无。解决了使用一个在线凝胶排阻吗色谱法(SEC)设置。SEC-SANS的可行性研究通过一系列的复杂和困难样本膜蛋白的嵌入nanodisc由磷脂和粒子蛋白质成分。SEC-SANS提供足够的数据这些系统的信噪比,同时同时规避聚合。结合SEC-SANS和SEC-SAXS数据,一个炼油结构的模型优化的依据研究了结构。

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