Lv Zhengjian, Banerjee Siddhartha, Zagorski Karen, Lyubchenko Yuri L
Department of Pharmaceutical Sciences, University of Nebraska Medical Center, Omaha, NE, USA.
Bruker Nano Surfaces Division, Santa Barbara, CA, USA.
Methods Mol Biol. 2018;1814:129-143. doi: 10.1007/978-1-4939-8591-3_8.
Nanoimaging methods, atomic force microscopy (AFM) in particular, are widely used to study the interaction of biological molecules with the supported lipid bilayer (SLB), which itself is a traditional model for cellular membranes. Success in these studies is based on the availability of a stable SLB for the required observation period, which can extend several hours. The application of AFM requires that the SLB have a smooth morphology, thus enabling visualization of proteins and other molecules on its surface. Herein, we describe protocols for SLB assembly by using 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (POPS) on a mica support. Our methodology enables us to assemble defect-free POPC and POPS SLBs that remain stable for at least 8 h. The application of such smooth and stable surfaces is illustrated by monitoring of the on-surface aggregation of amyloid proteins with the use of time-lapse AFM.
纳米成像方法，尤其是原子力显微镜（AFM），被广泛用于研究生物分子与支撑脂质双层（SLB）的相互作用，而支撑脂质双层本身就是细胞膜的传统模型。这些研究的成功基于在所需观察期内获得稳定的支撑脂质双层，观察期可能长达数小时。原子力显微镜的应用要求支撑脂质双层具有光滑的形态，从而能够在其表面可视化蛋白质和其他分子。在此，我们描述了在云母载体上使用1-棕榈酰-2-油酰-sn-甘油-3-磷酸胆碱（POPC）和1-棕榈酰-2-油酰-sn-甘油-3-磷酸-L-丝氨酸（POPS）组装支撑脂质双层的方案。我们的方法使我们能够组装无缺陷的POPC和POPS支撑脂质双层，它们至少能保持8小时的稳定性。通过使用延时原子力显微镜监测淀粉样蛋白在表面的聚集，说明了这种光滑且稳定表面的应用。
