Department of Chemistry, University of Guelph, Guelph, Ontario, N1G 2W1, Canada.
Langmuir. 2011 Aug 16;27(16):10072-87. doi: 10.1021/la201625c. Epub 2011 Jul 13.
Langmuir-Blodgett and Langmuir-Schaeffer methods were employed to deposit a mixed bilayer consisting of 90% of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and 10% of gramicidin (GD), a short 15 residue ion channel forming peptide, onto a Au(111) electrode surface. This architecture allowed us to investigate the effect of the electrostatic potential applied to the electrode on the orientation and conformation of DMPC molecules in the bilayer containing the ion channel. The charge density data were determined from chronocoulometry experiments. The electric field and the potential across the membrane were determined through the use of charge density curves. The magnitudes of potentials across the gold-supported biomimetic membrane were comparable to the transmembrane potential acting on a natural membrane. The information regarding the orientation and conformation of DMPC and GD molecules in the bilayer was obtained from photon polarization modulation infrared reflection absorption spectroscopy (PMIRRAS) measurements. The results show that the bilayer is adsorbed, in direct contact with the metal surface, when the potential across the interface is more positive than -0.4 V and is lifted from the gold surface when the potential across the interface is more negative than -0.4 V. This change in the state of the bilayer has a significant impact on the orientation and conformation of the phospholipid and gramicidin molecules. The potential induced changes in the membrane containing peptide were compared to the changes in the structure of the pure DMPC bilayer determined in earlier studies.
采用 Langmuir-Blodgett 和 Langmuir-Schaeffer 方法将由 90%的 1,2-二肉豆蔻酰-sn-甘油-3-磷酸胆碱(DMPC)和 10%的短 15 残基离子通道形成肽格兰菌素(GD)组成的混合双层沉积到 Au(111)电极表面。这种结构使我们能够研究施加到电极上的静电势对含有离子通道的双层中 DMPC 分子的取向和构象的影响。电荷密度数据是通过计时库仑法实验确定的。通过使用电荷密度曲线确定电场和膜两侧的电势。金支持的仿生膜两侧的电势幅度与作用于天然膜的跨膜电势相当。关于双层中 DMPC 和 GD 分子的取向和构象的信息是通过光子偏振调制红外反射吸收光谱(PMIRRAS)测量获得的。结果表明,当界面两侧的电势大于-0.4 V 时,双层以直接与金属表面接触的方式被吸附,而当界面两侧的电势小于-0.4 V 时,双层从金表面抬起。双层状态的这种变化对磷脂和格兰菌素分子的取向和构象有重大影响。将含有肽的膜的电位诱导变化与早期研究中确定的纯 DMPC 双层结构的变化进行了比较。