Melikishvili Sophie, Poturnayova Alexandra, Ionov Maksim, Bryszewska Maria, Vary Tomáš, Cirak Julius, Muñoz-Fernández María Ángeles, Gomez-Ramirez Rafael, de la Mata Francisco Javier, Hianik Tibor
Faculty of Mathematics, Physics and Informatics, Comenius University, Mlynska dolina, 842 48 Bratislava, Slovakia.
Faculty of Mathematics, Physics and Informatics, Comenius University, Mlynska dolina, 842 48 Bratislava, Slovakia; Institute of Animal Biochemistry and Genetics, Slovak Academy of Sciences, 900 28 Ivanka pri Dunaji, Slovakia.
Biochim Biophys Acta. 2016 Dec;1858(12):3005-3016. doi: 10.1016/j.bbamem.2016.09.005. Epub 2016 Sep 9.
In this study, dendrimers have been purposed as an alternative approach for delivery of HIV peptides to dendritic cells. We have investigated the interaction of dendriplexes formed from polyanionic HIV peptide Nef and cationic carbosilane dendrimer (CBD) with model lipid membranes - large unilamellar vesicles (LUVs), Langmuir monolayers and supported lipid membranes (sBLMs) containing various molar ratio of zwitterionic 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy (polyethylene glycol)-2000] (DSPE-PEG). In our experiments, the lipid membranes represented the model of the plasma membrane of the cell. PEGylated lipids were used in order to model glycocalyx which constitutes the outer leaflet of cellular membranes. The presence of PEGylated lipids resulted in an increase of the phase transition temperature of the lipid bilayer of LUVs, in a decrease of specific volume and adiabatic compressibility. Fluorescence anisotropy study suggests that PEGylated LUVs possessed higher lipid order and decreased fluidity when compared to zwitterionic DMPC vesicles. The interaction of dendriplexes with monolayers was accompanied by the formation of the aggregates as revealed by BAM experiments. This conclusion has been confirmed also by AFM imaging of sBLMs. We have demonstrated that dendriplexes interact with lipid membranes for all types of lipid composition. Moreover, the stronger interaction of cationic dendrimer/peptide complexes with lipid monolayers, vesicles and sBLMs was observed for membranes composed of zwitterionic lipids than for PEGylated lipid membranes. Increased concentration of PEGylated lipids made this interaction weaker.
在本研究中，树枝状大分子被用作将HIV肽递送至树突状细胞的一种替代方法。我们研究了由聚阴离子HIV肽Nef和阳离子碳硅烷树枝状大分子（CBD）形成的树枝状复合物与模型脂质膜——含有不同摩尔比两性离子1,2-二肉豆蔻酰-sn-甘油-3-磷酸胆碱（DMPC）和1,2-二硬脂酰-sn-甘油-3-磷酸乙醇胺-N-[甲氧基（聚乙二醇）-2000]（DSPE-PEG）的大单层囊泡（LUVs）、朗缪尔单层膜和支撑脂质膜（sBLMs）之间的相互作用。在我们的实验中，脂质膜代表细胞的质膜模型。使用聚乙二醇化脂质是为了模拟构成细胞膜外层的糖萼。聚乙二醇化脂质的存在导致LUVs脂质双层的相变温度升高，比容和绝热压缩性降低。荧光各向异性研究表明，与两性离子DMPC囊泡相比，聚乙二醇化LUVs具有更高的脂质有序度和更低的流动性。BAM实验表明，树枝状复合物与单层膜的相互作用伴随着聚集体的形成。sBLMs的原子力显微镜成像也证实了这一结论。我们已经证明，树枝状复合物与所有类型脂质组成的脂质膜都有相互作用。此外，观察到阳离子树枝状大分子/肽复合物与由两性离子脂质组成的膜的相互作用比与聚乙二醇化脂质膜的相互作用更强。聚乙二醇化脂质浓度的增加使这种相互作用减弱。
