Ayvazyan Naira M, Zaqaryan Naira A, Ghazaryan Narine A
Department of Biophysics, Faculty of Biology, Yerevan State University, Yerevan, Armenia.
Biochim Biophys Acta. 2012 May;1818(5):1359-64. doi: 10.1016/j.bbamem.2012.02.001. Epub 2012 Feb 8.
Studies on the interaction of snake venom and organized lipid interfaces have been conducted using a variety of systems, including BLMs, SUVs and GUVs. The present study was undertaken to elucidate how the plastic properties (namely, its microviscosity, thickness, permeability) of model membranes from native lipids of different tissues of rats change in the course of Macrovipera lebetina obtusa (MLO), Montivipera raddei (MR) and Naja kaouthia (NK) venoms processing. The presence of viper venom in organism leads to increasing of the electrical resistance of BLMs from liver and muscle lipids approximately on a sequence, while the BLMs from brain lipids have not shown noticeable differences of plastic properties compared to the control. Giant unilamellar vesicles (GUVs) with a mean diameter of 30μm have a minimum curvature and mimic cell membranes in this respect. Snake venom was added to the sample chamber before the vesicles were formed. The membrane fluorescence probes, ANS and pyrene, were used to assess the state of the membrane and specifically mark the phospholipid domains. Fluorescent spectra were acquired on a Varian fluorometer instrument. ANS and pyrene allow us to quantify the fluidity changes in the membrane by measuring of the fluorescence intensity. The presence of viper venom in GUVs media reveals a noticeable decreasing of membrane fluidity compared to the control, while the binding of fluorophores with GUVs modified by venom leads to the appearance of channel activity. These studies also emphasize the importance of a membrane surface curvature for its interaction with enzymatic components of venom.
人们已经使用包括脂质双分子层(BLMs)、小单层囊泡(SUVs)和大单层囊泡(GUVs)在内的多种系统,对蛇毒与有序脂质界面之间的相互作用进行了研究。本研究旨在阐明在钝尾草原蝰(MLO)、中介蝰(MR)和眼镜王蛇(NK)毒液作用过程中,大鼠不同组织天然脂质构成的模型膜的塑性特性(即其微粘度、厚度、通透性)如何变化。生物体内蝰蛇毒液的存在会使肝脏和肌肉脂质的BLMs电阻大致按顺序增加,而脑脂质的BLMs与对照组相比,塑性特性未显示出明显差异。平均直径为30μm的大单层囊泡(GUVs)具有最小曲率,在这方面模拟细胞膜。在囊泡形成之前,将蛇毒添加到样品池中。使用膜荧光探针1-苯胺基萘-8-磺酸(ANS)和芘来评估膜的状态,并特异性标记磷脂结构域。在瓦里安荧光计上获取荧光光谱。ANS和芘使我们能够通过测量荧光强度来量化膜中的流动性变化。与对照组相比,GUVs介质中蝰蛇毒液的存在显示出膜流动性明显降低,而荧光团与经毒液修饰的GUVs结合会导致通道活性的出现。这些研究还强调了膜表面曲率对其与毒液酶成分相互作用的重要性。