权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Development of integrated model biological membranes on solid substrates

固体基质上集成模型生物膜的开发

基本信息

批准号：
18510107
负责人：
MORIGAKI Kenichi
金额：
$ 2.58万
依托单位：
National Institute of Advanced Industrial Science and Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

Micropatterned phospholipid bilayers on solid substrates offer an attractive platform for various applications, such as high throughput drug screening. We are currently developing a method for generating micropatterned bilayers composed of polymerized and fluid lipid bilayers. Lithographic photopolymerization of a diacetylene-containing phospholipid (DiynePC) allowed facile fabrication of compartmentalized arrays of fluid lipid membranes. In the present project, we could find a key experimental parameter, that significantly influenced the homogeneity and quality of the fabricated polymeric bilayers, namely the temperature at which monolayers of monomeric DiynePC were formed on the water surface and transferred onto solid substrates by the Langmuir-Blodgett/Langmuir-Schaefer (LB/LS) technique. Using the fluorescence microscopy and atomic force microscopy, it was found that polymerized bilayers were homogeneous, if bilayers of DiynePC were prepared below the triple point temperature (ca. 20℃) of the monolayer, where a direct transition from the gaseous state to the liquid condensed state occurred. The differences were attributed to the domain structures in the monolayer that was transferred from the water surface to the substrate. Another important result from the project is the development of a method to incorporate lipid membranes with membrane proteins. We used mixtures of short and long chain phospholipid, 1, 2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC). We demonstrated that formation of planar bilayers was significantly accelerated in the presence of DHPC. Furthermore, membranes containing calcium channels(SR membranes from rabbit muscle) could be incorporated in the model membrane. These results suggest that the use of the mixtures of short and long chain phospholipids could be a convenient means for preparing integrated model membranes.

固体基质上的微图案化磷脂双层为各种应用提供了有吸引力的平台，例如高通量药物筛选。我们目前正在开发一种方法，用于产生微图案化的双层聚合和流体脂质双层组成。含二乙炔的磷脂（DiynePC）的光刻光聚合允许流体脂质膜的区室化阵列的容易制造。在本项目中，我们可以找到一个关键的实验参数，显着影响制造的聚合物双层的均匀性和质量，即单体DiynePC的单层在水表面上形成的温度和转移到固体基质上的LB/LS技术。使用荧光显微镜和原子力显微镜，发现聚合的双层是均匀的，如果DiynePC的双层在低于三相点温度（约100 ℃）制备，则聚合的双层是均匀的。20℃）时，发生了从气态到液态凝聚态的直接转变。这种差异归因于从水表面转移到基底的单分子膜中的畴结构。该项目的另一个重要成果是开发了一种将脂质膜与膜蛋白结合的方法。我们使用短链和长链磷脂的混合物，1，2-二己酰基-sn-甘油基-3-磷酸胆碱（DHPC）和1-棕榈酰基-2-油酰基-sn-甘油基-3-磷酸胆碱（POPC）。我们证明，平面双层的形成显着加速DHPC的存在下。此外，含有钙通道的膜（来自兔肌肉的SR膜）可以并入模型膜中。这些结果表明，使用的短链和长链磷脂的混合物可以是一种方便的手段，用于制备集成的模型膜。