Institute of Analytical Chemistry of the Czech Academy of Sciences, v. v. i., Veveří 97, 602 00 Brno, Czech Republic.
Department of Chemistry, University of Helsinki, A.I. Virtasen aukio 1, P.O. Box 55, FI-00014, Finland.
Biochim Biophys Acta Biomembr. 2020 Feb 1;1862(2):183115. doi: 10.1016/j.bbamem.2019.183115. Epub 2019 Nov 5.
The cell membrane is mainly composed of lipid bilayers with inserted proteins and carbohydrates. Lipid bilayers made of purified or synthetic lipids are widely used for estimating the effect of target compounds on cell membranes. However, the composition of such biomimetic membranes is much simpler than the composition of biological membranes. Interactions between compounds and simple composition biomimetic membranes might not demonstrate the effect of target compounds as precisely as membranes with compositions close to real organisms. Therefore, the aim of our study is to construct biomimetic membrane closely mimicking the state of natural membranes. Liposomes were prepared from lipids extracted from L-α-phosphatidylcholine, Escherichia coli, yeast (Saccharomyces cerevisiae) and bovine liver cells through agitation and sonication. They were immobilized onto silicon dioxide (SiO) sensor surfaces using N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid buffer with calcium chloride. The biomimetic membranes were successfully immobilized onto the SiO sensor surface and detected by nanoplasmonic sensing. The immobilized membranes were exposed to choline carboxylates. The membrane disruption effect was, as expected, more pronounced with increasing carbohydrate chain length of the carboxylates. The results correlated with the toxicity values determined using Vibrio fischeri bacteria. The yeast extracted lipid membranes had the strongest response to introduction of choline laurate while the bovine liver lipid extracted liposomes were the most sensitive towards the shorter choline carboxylates. This implies that the composition of the cell membrane plays a crucial role upon interaction with choline carboxylates, and underlines the necessity of testing membrane systems of different origin to obtain an overall image of such interactions.
细胞膜主要由双层脂类和插入的蛋白质和碳水化合物组成。由纯化或合成脂质制成的双层脂类广泛用于估计目标化合物对细胞膜的影响。然而,这种仿生膜的组成比生物膜的组成简单得多。化合物与简单组成的仿生膜之间的相互作用可能不如组成接近真实生物体的膜那样精确地显示目标化合物的作用。因此,我们研究的目的是构建尽可能模拟天然膜状态的仿生膜。通过搅拌和超声处理,从 L-α-磷脂酰胆碱、大肠杆菌、酵母(酿酒酵母)和牛肝细胞中提取的脂质制备脂质体。它们通过使用含有氯化钙的 N-(2-羟乙基)哌嗪-N'-2-乙磺酸缓冲液固定在二氧化硅(SiO)传感器表面上。仿生膜成功地固定在 SiO 传感器表面上,并通过纳米等离子体传感进行检测。将固定化膜暴露于胆碱羧酸酯。正如预期的那样,随着羧酸酯的碳水化合物链长的增加,膜的破坏效应更为明显。结果与使用发光菌(Vibrio fischeri)细菌确定的毒性值相关。酵母提取的脂质膜对引入月桂酰胆碱的反应最强,而牛肝脂质提取的脂质体对较短的胆碱羧酸酯最敏感。这意味着细胞膜的组成在与胆碱羧酸酯相互作用时起着至关重要的作用,并强调了测试不同来源的膜系统以获得此类相互作用的整体图像的必要性。