Kostelic Marius M, Zak Ciara K, Jayasekera Hiruni S, Marty Michael T
Department of Chemistry and Biochemistry, University of Arizona, Tucson, Arizona 85721, United States.
Bio5 Institute, University of Arizona, Tucson, Arizona 85721, United States.
Anal Chem. 2021 Apr 13;93(14):5972-5979. doi: 10.1021/acs.analchem.1c00735. Epub 2021 Apr 2.
Native mass spectrometry (MS) with nanodiscs is a promising technique for characterizing membrane protein and peptide interactions in lipid bilayers. However, prior studies have used nanodiscs made of only one or two lipids, which lack the complexity of a natural lipid bilayer. To better model specific biological membranes, we developed model mammalian, bacterial, and mitochondrial nanodiscs with up to four different phospholipids. Careful selection of lipids with similar masses that balance the fluidity and curvature enabled these complex nanodiscs to be assembled and resolved with native MS. We then applied this approach to characterize the specificity and incorporation of LL-37, a human antimicrobial peptide, in single-lipid nanodiscs versus model bacterial nanodiscs. Overall, development of these model membrane nanodiscs reveals new insights into the assembly of complex nanodiscs and provides a useful toolkit for studying membrane protein, peptide, and lipid interactions in model biological membranes.
使用纳米圆盘的原生质谱（MS）是一种用于表征脂质双层中膜蛋白和肽相互作用的有前途的技术。然而，先前的研究使用的是仅由一两种脂质制成的纳米圆盘，它们缺乏天然脂质双层的复杂性。为了更好地模拟特定的生物膜，我们开发了含有多达四种不同磷脂的模型哺乳动物、细菌和线粒体纳米圆盘。仔细选择具有相似质量且能平衡流动性和曲率的脂质，使得这些复杂的纳米圆盘能够通过原生质谱进行组装和解析。然后，我们应用这种方法来表征人抗菌肽LL-37在单脂质纳米圆盘与模型细菌纳米圆盘中的特异性和掺入情况。总体而言，这些模型膜纳米圆盘的开发揭示了对复杂纳米圆盘组装的新见解，并为研究模型生物膜中的膜蛋白、肽和脂质相互作用提供了一个有用的工具包。
