Department of Chemistry and Biochemistry, University of Arizona, Tucson, AZ 85721, United States.
Bio5 Institute, University of Arizona, Tucson, AZ 85721, United States.
J Am Soc Mass Spectrom. 2023 Sep 6;34(9):2006-2015. doi: 10.1021/jasms.3c00170. Epub 2023 Jul 31.
Lipids play critical roles in modulating membrane protein structure, interactions, and activity. Nanodiscs provide a tunable membrane mimetic that can model these endogenous protein-lipid interactions in a nanoscale lipid bilayer. However, most studies of membrane proteins with nanodiscs use simple synthetic lipids that lack the headgroup and fatty acyl diversity of natural extracts. Prior research has successfully used natural lipid extracts in nanodiscs that more accurately mimic natural environments, but it is not clear how nanodisc assembly may bias the incorporated lipid profiles. Here, we applied lipidomics to investigate how nanodisc assembly conditions affect the profile of natural lipids in nanodiscs. Specifically, we tested the effects of assembly temperature, nanodisc size, and lipidome extract complexity. Globally, our analysis demonstrates that the lipids profiles are largely unaffected by nanodisc assembly conditions. However, a few notable changes emerged within individual lipids and lipid classes, such as a differential incorporation of cardiolipin and phosphatidylglycerol lipids from the polar lipid extract at different temperatures. Conversely, some classes of brain lipids were affected by nanodisc size at higher temperatures. Collectively, these data enable the application of nanodiscs to study protein-lipid interactions in complex lipid environments.
脂质在调节膜蛋白结构、相互作用和活性方面起着至关重要的作用。纳米盘提供了一种可调节的膜模拟物，可以在纳米尺度的脂质双层中模拟这些内源性的蛋白-脂质相互作用。然而，大多数使用纳米盘研究膜蛋白的研究都使用简单的合成脂质，这些脂质缺乏天然提取物的头部基团和脂肪酸多样性。先前的研究已经成功地在纳米盘中使用了天然脂质提取物，以更准确地模拟自然环境，但目前尚不清楚纳米盘组装如何影响所包含的脂质谱。在这里，我们应用脂质组学来研究纳米盘组装条件如何影响纳米盘中天然脂质的分布。具体来说，我们测试了组装温度、纳米盘尺寸和脂质组提取物复杂性的影响。总的来说，我们的分析表明，脂质分布在很大程度上不受纳米盘组装条件的影响。然而，在个别脂质和脂质类中出现了一些值得注意的变化，例如在不同温度下，极性脂质提取物中的心磷脂和磷脂酰甘油脂质的不同掺入。相反，一些类别的脑脂质在较高温度下受到纳米盘尺寸的影响。总的来说，这些数据使得纳米盘能够应用于研究复杂脂质环境中的蛋白-脂质相互作用。
