β-辛基葡萄糖苷和磷酸盐溶液对单油酸甘油酯中间相相行为的影响。

Effects of detergent β-octylglucoside and phosphate salt solutions on phase behavior of monoolein mesophases.

机构信息

Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana-Champaign, Urbana, Illinois.

出版信息

Biophys J. 2013 Oct 15;105(8):1848-59. doi: 10.1016/j.bpj.2013.09.009.

Abstract

Using small-angle x-ray scattering (SAXS), we investigated the phase behavior of mesophases of monoolein (MO) mixed with additives commonly used for the crystallization of membrane proteins from lipidic mesophases. In particular, we examined the effect of sodium and potassium phosphate salts and the detergent β-octylglucoside (βOG) over a wide range of compositions relevant for the crystallization of membrane proteins in lipidic mesophases. We studied two types of systems: 1), ternary mixtures of MO with salt solutions above the hydration boundary; and 2), quaternary mixtures of MO with βOG and salt solutions over a wide range of hydration conditions. All quaternary mixtures showed highly regular lyotropic phase behavior with the same sequence of phases (Lα, Ia3d, and Pn3m) as MO/water mixtures at similar temperatures. The effects of additives in quaternary systems agreed qualitatively with those found in ternary mixtures in which only one additive is present. However, quantitative differences in the effects of additives on the lattice parameters of fully hydrated mesophases were found between ternary and quaternary mixtures. We discuss the implications of these findings for mechanistic investigations of membrane protein crystallization in lipidic mesophases and for studies of the suitability of precipitants for mesophase-based crystallization methods.

摘要

利用小角 X 射线散射(SAXS),我们研究了单油酸甘油酯(MO)与脂质相中常用的膜蛋白结晶添加剂混合形成的中间相的相行为。特别是,我们考察了在脂质相中的膜蛋白结晶相关的广泛组成范围内,钠盐和钾盐以及去污剂β-辛基葡萄糖苷(βOG)对中间相的影响。我们研究了两种类型的体系:1)MO 与盐溶液的三元混合物,在水合边界之上;以及 2)MO 与βOG 和盐溶液的四元混合物,在广泛的水合条件下。所有的四元混合物都表现出高度规则的溶致相行为,在相似的温度下具有与 MO/水混合物相同的相序列(Lα、Ia3d 和 Pn3m)。在四元体系中,添加剂的影响在定性上与仅存在一种添加剂的三元混合物中的影响一致。然而,在完全水合中间相的晶格参数方面,添加剂对三元和四元混合物的影响存在定量差异。我们讨论了这些发现对脂质相中的膜蛋白结晶的机制研究以及对沉淀剂在基于中间相的结晶方法中的适用性研究的意义。

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