Detergent-soluble proteins (DSPs) are commonly dissolved in lipid buffers for NMR experiments, but the huge lipid proton signal prevents recording of high-quality spectra. The use of costly deuterated lipids is thus required to replace nondeuterated ones. With conventional methods, detergents like dodecylphosphocholine (DPC) cannot be fully exchanged due to their high binding affinity to hydrophobic proteins. We propose an original and simple protocol which combines the use of acetonitrile, dialysis and lyophilization to disrupt the binding of lipids to the protein and allow their indirect replacement by their deuterated equivalents, while maintaining the native structure of the protein. Moreover, by this protocol, the detergent-to-protein molar ratio can be controlled as it challenges the protein structure. This protocol was applied to solubilize the Vpx protein that was followed upon addition of DPC-d₃₈ by ¹H-¹⁵N SOFAST-HMQC spectra and the best detergent-to-DSPs molar ratio was obtained for structural studies.

中文翻译：

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乙腈允许用氘化脂质去污剂间接替代非氘化脂质去污剂，用于去污剂可溶性蛋白质的核磁共振研究

去污剂可溶性蛋白质 (DSP) 通常溶解在用于 NMR 实验的脂质缓冲液中，但巨大的脂质质子信号阻碍了高质量光谱的记录。因此需要使用昂贵的氘代脂质来代替非氘代脂质。使用传统方法，十二烷基磷酸胆碱 (DPC) 等去污剂由于对疏水蛋白的高结合亲和力而不能完全交换。我们提出了一种原始且简单的方案，该方案结合使用乙腈、透析和冻干来破坏脂质与蛋白质的结合，并允许它们被氘代等价物间接替代，同时保持蛋白质的天然结构。此外，通过该协议，可以控制洗涤剂与蛋白质的摩尔比，因为它挑战了蛋白质结构。d ₃₈ × ¹ H- ¹⁵ N SOFAST-HMQC 光谱和最佳去污剂与 DSP 的摩尔比用于结构研究。