Okazaki Takashi, Inaba Takehiko, Tatsu Yoshiro, Tero Ryugo, Urisu Tsuneo, Morigaki Kenichi
Research Institute for Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Ikeda 563-8577, Japan.
Langmuir. 2009 Jan 6;25(1):345-51. doi: 10.1021/la802670t.
Substrate supported planar lipid bilayers (SPBs) are versatile models of the biological membrane in biophysical studies and biomedical applications. We previously developed a methodology for generating SPBs composed of polymeric and fluid phospholipid bilayers by using a photopolymerizable diacetylene phospholipid (DiynePC). Polymeric bilayers could be generated with micropatterns by conventional photolithography, and the degree of polymerization could be controlled by modulating UV irradiation doses. After removing nonreacted monomers, fluid lipid membranes could be integrated with polymeric bilayers. Herein, we report on a quantitative study of the morphology of polymeric bilayer domains and their obstruction toward lateral diffusion of membrane-associated molecules. Atomic force microscopy (AFM) observations revealed that polymerized DiynePC bilayers were formed as nanometer-sized domains. The ratio of polymeric and fluid bilayers could be modulated quantitatively by changing the UV irradiation dose for photopolymerization. Lateral diffusion coefficients of lipid molecules in fluid bilayers were measured by fluorescence recovery after photobleaching (FRAP) and correlated with the amount of polymeric bilayer domains on the substrate. Controlled domain structures, lipid compositions, and lateral mobility in the model membranes should allow us to fabricate model membranes that mimic complex features of biological membranes with well-defined structures and physicochemical properties.
在生物物理研究和生物医学应用中,底物支撑的平面脂质双层膜(SPBs)是生物膜的通用模型。我们之前开发了一种方法,通过使用可光聚合的二乙炔磷脂(DiynePC)来生成由聚合物和流体磷脂双层组成的SPBs。聚合物双层可以通过传统光刻技术形成微图案,并且可以通过调节紫外线照射剂量来控制聚合度。去除未反应的单体后,流体脂质膜可以与聚合物双层整合。在此,我们报告了对聚合物双层结构域形态及其对膜相关分子横向扩散的阻碍的定量研究。原子力显微镜(AFM)观察表明,聚合的DiynePC双层形成为纳米级结构域。通过改变用于光聚合的紫外线照射剂量,可以定量调节聚合物双层和流体双层的比例。通过光漂白后荧光恢复(FRAP)测量流体双层中脂质分子的横向扩散系数,并将其与底物上聚合物双层结构域的数量相关联。模型膜中可控的结构域结构、脂质组成和横向流动性应使我们能够制造出模仿具有明确结构和物理化学性质的生物膜复杂特征的模型膜。