Lotta T I, Laakkonen L J, Virtanen J A, Kinnunen P K
Department of Membrane Physics, KSV Research Laboratories, Helsinki, Finland.
Chem Phys Lipids. 1988 Jan;46(1):1-12. doi: 10.1016/0009-3084(88)90107-7.
Monomolecular films of 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC) and 1-palmitoyl-2-[10-(pyren-1-yl)decanoyl]-sn-glycero-3-phosphatidylc holine (PPDPC) were transferred from an air/water interface onto a germanium attenuated total reflection crystal by the Langmuir-Blodgett (LB) technique. The assemblies were thereafter investigated by Fourier transform infrared-attenuated total reflection (FTIR-ATR) spectroscopy. To determine the molecular organization in the deposited layers we monitored the CH2 and C = O stretching and the CH2 bending regions of the infrared spectra of these lipids in detail. Using Fourier self-deconvolution technique, the carbonyl stretching mode was resolved into two models corresponding to the conformational differences in the ester linkages of the phospholipid sn-1 and sn-2 acyl chains. By varying the temperature of the subphase and using different surface pressures, we were able to transfer different conformational states of DPPC onto a germanium ATR crystal. Deposition of DPPC at 40 mN m-1 and at 15 degrees C or at 20 mN m-1 and at 35 degrees C results in LB-assemblies in ordered or disordered states, respectively, as judged by the IR spectra. These structures in LB films correspond to the state of DPPC in liposomes below and above the temperature of the order-disorder phase transition. Irrespective of the surface pressure and subphase temperature used during the deposition, an ordering process was found in DPPC films when the number of the transferred layers was increased from one to five. The pyrene-labelled phosphatidylcholine analogue, PPDPC, behaved differently from DPPC. In the case where one to three layers of PPDPC transferred at 35 mN m-1 and at 20 degrees C only conformational structures resembling those in fully hydrated liposomes above the main transition temperature were observed.
通过朗缪尔-布洛杰特(LB)技术,将1,2-二棕榈酰-sn-甘油-3-磷酸胆碱(DPPC)和1-棕榈酰-2-[10-(芘-1-基)癸酰]-sn-甘油-3-磷酸胆碱(PPDPC)的单分子膜从气/水界面转移到锗衰减全反射晶体上。之后,通过傅里叶变换红外衰减全反射(FTIR-ATR)光谱对这些组装体进行研究。为了确定沉积层中的分子排列,我们详细监测了这些脂质红外光谱中的CH2和C=O伸缩振动以及CH2弯曲振动区域。使用傅里叶自去卷积技术,羰基伸缩振动模式被解析为两种模式,分别对应磷脂sn-1和sn-2酰基链酯键中的构象差异。通过改变亚相温度并使用不同的表面压力,我们能够将DPPC的不同构象状态转移到锗ATR晶体上。根据红外光谱判断,在40 mN m-1和15℃或20 mN m-1和35℃下沉积DPPC分别会形成有序或无序状态的LB组装体。这些LB膜中的结构对应于脂质体中DPPC在有序-无序相变温度以下和以上的状态。无论在沉积过程中使用的表面压力和亚相温度如何,当转移层数从一层增加到五层时,在DPPC膜中都发现了一个有序化过程。芘标记的磷脂酰胆碱类似物PPDPC的行为与DPPC不同。在35 mN m-1和20℃下转移一到三层PPDPC的情况下,仅观察到类似于主要转变温度以上完全水合脂质体中的构象结构。