Picquart M, Nicolas E, Lavialle F
Laboratoire de Physique Moléculaire et Biologique, CNRS, Université René Descartes, Paris, France.
Eur Biophys J. 1989;17(3):143-9. doi: 10.1007/BF00254768.
Perturbations induced by a toxic lectin (ricin) on lipid organisation of model membranes prepared with DPPC and DPPC-cerebrosides mixtures have been analysed by Raman and infrared spectroscopy, two powerful and non-invasive methods. Our approach involves the observation of changes in the vibrational spectra of liquid multilayers in the PO2-, C = O and CH2 spectral regions for two lipid:ricin molar ratios (225:1, 75:1). The interfacial and polar regions of the multilayers, analysed by FTIR, appear to be perturbed by the protein. With both kinds of membranes, ricin mainly perturbs the C = O ester groups of the sn-2 acylchain of DPPC. In the PO2- stretching region, the frequency shifts are correlated with changes in polar group hydration. In the hydrophobic core of the multilayer membrane studied by Raman spectroscopy, the interaction of ricin is associated with changes in lipid packing. These perturbations depend upon the lipid composition of the membrane. With DPPC membranes, an affect is detected at temperatures lower than Tm. It corresponds to a decrease of the lipid ordering. With DPPC-cer membranes, the protein increases the acyl-chain packing order regardless of the temperature of the experiments (10 degrees C less than T less than 75 degrees C). No perturbation of Tm is observed after addition of ricin to either DPPC or DPPC-cer membranes. The different perturbations detected by Raman and FTIR suggest that ricin mainly interacts with the interfacial domains of the membranes.
通过拉曼光谱和红外光谱这两种强大的非侵入性方法,分析了一种有毒凝集素(蓖麻毒素)对用二棕榈酰磷脂酰胆碱(DPPC)和DPPC - 脑苷脂混合物制备的模型膜脂质组织的扰动。我们的方法包括观察两种脂质与蓖麻毒素摩尔比(225:1、75:1)下,PO₂⁻、C = O和CH₂光谱区域中液体多层膜振动光谱的变化。通过傅里叶变换红外光谱(FTIR)分析的多层膜的界面和极性区域似乎受到了蛋白质的扰动。对于这两种膜,蓖麻毒素主要扰动DPPC的sn - 2酰基链的C = O酯基。在PO₂⁻伸缩区域,频率变化与极性基团水合作用的变化相关。在用拉曼光谱研究的多层膜疏水核心中,蓖麻毒素的相互作用与脂质堆积的变化有关。这些扰动取决于膜的脂质组成。对于DPPC膜,在低于熔点温度(Tm)时检测到影响。这对应于脂质有序性的降低。对于DPPC - 脑苷脂膜(DPPC - cer),无论实验温度如何(10℃<T<75℃),蛋白质都会增加酰基链的堆积顺序。向DPPC或DPPC - cer膜中添加蓖麻毒素后,未观察到Tm的扰动。拉曼光谱和FTIR检测到的不同扰动表明,蓖麻毒素主要与膜的界面区域相互作用。