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Membrane proteins in magnetically aligned phospholipid polymer discs for solid-state NMR spectroscopy.
Biochimica et Biophysica Acta (BBA) - Biomembranes ( IF 2.8 ) Pub Date : 2020-05-01 , DOI: 10.1016/j.bbamem.2020.183333
Sang Ho Park 1 , Jiaqian Wu 1 , Yong Yao 2 , Chandan Singh 2 , Ye Tian 2 , Francesca M Marassi 2 , Stanley J Opella 1
Affiliation  

Well-hydrated phospholipid bilayers provide a near-native environment for membrane proteins. They enable the preparation of chemically-defined samples suitable for NMR and other spectroscopic experiments that reveal the structure, dynamics, and functional interactions of the proteins at atomic resolution. The synthetic polymer styrene maleic acid (SMA) can be used to prepare detergent-free samples that form macrodiscs with diameters greater than 30 nm at room temperature, and spontaneously align in the magnetic field of an NMR spectrometer at temperatures above 35 °C. Here we show that magnetically aligned macrodiscs are particularly well suited for solid-state NMR experiments of membrane proteins because the SMA-lipid assembly both immobilizes the embedded protein and provides uniaxial order for oriented sample (OS) solid-state NMR studies. We show that aligned macrodiscs incorporating four different membrane proteins with a wide range of sizes and topological complexity yield high-resolution OS solid-state NMR spectra. The work is dedicated to Michelle Auger who made key contributions to the field of membrane and membrane protein biophysics.

中文翻译:

用于固态核磁共振光谱的磁性排列的磷脂聚合物盘中的膜蛋白。

水合良好的磷脂双层为膜蛋白提供了接近天然的环境。它们能够制备适合 NMR 和其他光谱实验的化学定义样品,以原子分辨率揭示蛋白质的结构、动力学和功能相互作用。合成聚合物苯乙烯马来酸 (SMA) 可用于制备不含洗涤剂的样品,这些样品在室温下形成直径大于 30 nm 的大圆盘,并在 35 °C 以上的温度下在 NMR 光谱仪的磁场中自发排列。在这里,我们表明磁性对齐的宏圆盘特别适合膜蛋白的固态 NMR 实验,因为 SMA 脂质组件既固定嵌入的蛋白质,又为定向样品 (OS) 固态 NMR 研究提供单轴顺序。我们表明,对齐的宏盘结合了四种不同的膜蛋白,具有广泛的尺寸和拓扑复杂性,可产生高分辨率的 OS 固态 NMR 光谱。这项工作献给在膜和膜蛋白生物物理学领域做出重要贡献的米歇尔·奥格 (Michelle Auger)。
更新日期:2020-05-01
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