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Solution NMR of proteins within polyacrylamide gels : diffusional properties and residual alignment by mechanical stress or embedding of oriented purple membranes

机译:聚丙烯酰胺凝胶中蛋白质的溶液核磁共振:扩散特性和机械应力的残余排列或定向紫膜的嵌入

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摘要

The diffusive properties of biomacromolecules within the aqueous phase of polyacrylamide gels are described. High quality NMR spectra can be obtained under such conditions. As compared to water, a fivefold reduction in the translational diffusion constant, but only a 1.6-fold decrease (1.4-fold increase) in amide-15N T2 (T1) are observed for human ubiquitin within a 10% acrylamide gel. Weak alignment of the solute macromolecules can be achieved within such gels by vertical or radial compression or by the embedding of magnetically oriented purple membrane fragments. The methods are applied to deriveresidual dipolar couplings for human HIV-1 Nef and ubiquitin.
机译:描述了生物大分子在聚丙烯酰胺凝胶水相内的扩散特性。在这样的条件下可以获得高质量的NMR光谱。与水相比,在10%丙烯酰胺凝胶中的人泛素中酰胺15N T2(T1)的翻译扩散常数降低了5倍,但仅降低了1.6倍(增加了1.4倍)。溶质大分子的弱排列可在此类凝胶中通过垂直或径向压缩或通过嵌入磁性取向的紫色膜片段来实现。该方法用于推导人HIV-1 Nef和泛素的残留偶极偶联。

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