Nielsen Søren B, Otzen Daniel E
Arla Foods Ingredients Group P/S, Technology and Functionality, R&D, Protein Chemistry & Functionality, Videbæk, Denmark.
Department of Molecular Biology and Genetics, Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Aarhus C, Denmark.
Methods Mol Biol. 2019;2003:31-52. doi: 10.1007/978-1-4939-9512-7_2.
Extensive studies on the spontaneous collapse of phospholipid vesicles into supported lipid bilayers (SLBs) have led to procedures which allow SLB formation on a wealth of substrates and lipid compositions. SLBs provide a widely accepted and versatile model system which mimics the natural cell membrane separating the extracellular and intracellular fluids of the living cell. The quartz crystal microbalance with dissipation monitoring (QCM-D) has been central in both the understanding of vesicle collapse into SLBs on various substrates but also in probing the kinetics and mechanisms of biomolecular interactions with SLBs in real time. We describe a robust procedure to form SLBs of zwitterionic and charged lipids on SiO sensor crystals which subsequently can be exploited to probe the interaction between proteins and peptides with the SLB.
对磷脂囊泡自发塌陷形成支撑脂质双层(SLB)的广泛研究催生了一系列方法,这些方法能使SLB在多种底物和脂质组成上形成。SLB提供了一个被广泛接受且通用的模型系统,它模拟了分隔活细胞细胞外液和细胞内液的天然细胞膜。具有耗散监测功能的石英晶体微天平(QCM-D)在理解囊泡在各种底物上塌陷形成SLB的过程中发挥了核心作用,同时也能实时探测生物分子与SLB相互作用的动力学和机制。我们描述了一种在SiO传感器晶体上形成两性离子和带电脂质的SLB的可靠方法,随后可利用该方法探测蛋白质和肽与SLB之间的相互作用。