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Kinetically Stable Triglyceride-Based Nanodroplets and Their Interactions with Lipid-Specific Proteins
Langmuir ( IF 3.7 ) Pub Date : 2018-07-07 00:00:00 , DOI: 10.1021/acs.langmuir.8b02180
Valerija Vezočnik 1 , Vesna Hodnik 1 , Simona Sitar 2 , Halil I. Okur 3 , Magda Tušek-Žnidarič 4 , Cornelis Lütgebaucks 3 , Kristina Sepčić 1 , Ksenija Kogej 5 , Sylvie Roke 3 , Ema Žagar 2 , Peter Maček 1
Affiliation  

Understanding of the interactions between proteins and natural and artificially prepared lipid membrane surfaces and embedded nonpolar cores is important in studies of physiological processes and their pathologies and is applicable to nanotechnologies. In particular, rapidly growing interest in cellular droplets defines the need for simplified biomimetic lipid model systems to overcome in vivo complexity and variability. We present a protocol for the preparation of kinetically stable nanoemulsions with nanodroplets composed of sphingomyelin (SM) and cholesterol (Chol), as amphiphilic surfactants, and trioleoylglycerol (TOG), at various molar ratios. To prepare stable SM/Chol-coated monodisperse lipid nanodroplets, we modified a reverse phase evaporation method and combined it with ultrasonication. Lipid composition, ζ-potential, gyration and hydrodynamic radius, shape, and temporal stability of the lipid nanodroplets were characterized and compared to extruded SM/Chol large unilamellar vesicles. Lipid nanodroplets and large unilamellar vesicles with theoretical SM/Chol/TOG molar ratios of 1/1/4.7 and 4/1/11.7 were further investigated for the orientational order of their interfacial water molecules using a second harmonic scattering technique, and for interactions with the SM-binding and Chol-binding pore-forming toxins equinatoxin II and perfringolysin O, respectively. The surface characteristics (ζ-potential, orientational order of interfacial water molecules) and binding of these proteins to the nanodroplet SM/Chol monolayers were similar to those for the SM/Chol bilayers of the large unilamellar vesicles and SM/Chol Langmuir monolayers, in terms of their surface structures. We propose that such SM/Chol/TOG nanoparticles with the required lipid compositions can serve as experimental models for monolayer membrane to provide a system that imitates the natural lipid droplets.

中文翻译:

运动稳定的甘油三酸酯基纳米滴及其与脂质特异性蛋白的相互作用。

了解蛋白质与天然和人工制备的脂质膜表面以及嵌入的非极性核心之间的相互作用在生理过程及其病理学研究中很重要,并且适用于纳米技术。特别地,对细胞液滴的快速增长的兴趣定义了对简化的仿生脂质模型系统的需要,以克服体内的复杂性和可变性。我们提出了一种制备具有动力学稳定性的纳米乳剂的协议,该纳米乳剂由鞘磷脂(SM)和胆固醇(Chol)作为两亲性表面活性剂和三油酰甘油(TOG)以不同的摩尔比组成。为了制备稳定的SM / Chol包被的单分散脂质纳米滴,我们改进了反相蒸发方法,并将其与超声处理相结合。脂质成分ζ电位 表征了脂质纳米滴的回转和流体动力学半径,形状和时间稳定性,并与挤压的SM / Chol大单层囊泡进行了比较。使用二次谐波散射技术进一步研究了脂质纳米滴和理论SM / Chol / TOG摩尔比为1/1 / 4.7和4/1 / 11.7的大单层囊泡的界面水分子的取向顺序,以及与SM结合和Chol结合的成孔毒素马毒素II和穿孔菌溶血素O。这些蛋白质的表面特性(ζ势,界面水分子的取向顺序)以及与纳米液滴SM / Chol单层的结合与大型单层囊泡的SM / Chol双层和SM / Chol Langmuir单层的相似,就其表面结构而言。我们建议具有所需脂质组成的此类SM / Chol / TOG纳米颗粒可以用作单层膜的实验模型,以提供模拟天然脂质液滴的系统。
更新日期:2018-07-07
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