A Self-assembly Route for Double Bilayer Lipid Membrane Formation
A Self-assembly Route for Double Bilayer Lipid Membrane Formation
复制标题
DOI：
10.1002/cphc.200900798
复制
发表时间：
2010-02-22
期刊：
影响因子：
2.9
通讯作者：
Evans, Stephen D.
中科院分区：
文献类型：
--
作者：
Han, Xiaojun;Achalkumar, Ammathnadu S.;Evans, Stephen D.
固体支持的脂质膜提供了一个简单的仿生模型系统，适用于研究广泛的膜相关现象，重要的是允许使用许多表面分析技术[1-8]，从AFM到阻抗谱，用于表征这些过程。此外，还认为这样的系统可以应用于药物筛选、生物传感或蛋白质分离/结晶。[9]然而，在自然界中，存在许多双双层天然存在的情况，例如在线粒体[10]或在间隙连接处跨越两个脂质双层的复合物[11]中，并且由于这个原因，期望能够产生双双层模拟物作为支持的双层场的延伸。以前已经证明可以使用Langmuir-Blodgett技术创建这样的结构，[12]然而，这具有与跨膜蛋白的掺入不相容的缺点，并且膜是通过空气-水界面转移而创建的。因此，基于囊泡自组装的方法将在双双层可用于的应用类型方面提供重大进展。Murray等人最近已经证明了一种先前观察到的现象，即第二个双层可以组装在链霉亲和素蛋白膜的顶部，附着在第一个双层上。[13]此外，Chung等人还表明，巨大单层囊泡（GUV）破裂可导致使用互补DNA序列拴系到第一双层的第二双层的形成。[14]类似地，Kaizuka和格罗夫斯已经报道了GUV在脂质双层上破裂以形成模型膜间连接;[15]并且Tabaei等人已经证明了一种方法，其中DNA双链体用于将多个盘形脂质“双胞”拴系到双层上。[16]虽然这些系统不能直接用于研究双双层现象，但它们表明通过自组装实现双双层的原理是可行的。在这里，我们描述了一种使用NHS/EDC [17]化学[羟基-2，5-二氧代吡咯烷-3-磺酸钠盐（NHS）和N-乙基-N ′-（3-二甲氨基丙基）碳二亚胺盐酸盐（EDC）]在固体支持物上形成双双层脂质膜（dBLMs）的新方法。
Solid supported lipid membranes provide a simple biomimetic model system that is suitable for studying a wide range of membrane related phenomena and importantly permits the use of a number of surface analytical techniques [1–8] from AFM to impedance spectroscopy to be used in characterising such processes. Additionally, it is also believed that such systems could find application for drug screening, biosensing or in protein separation/crystallisation.[9] In nature there are however a number of situations in which double bilayers naturally occur, for example in mitochondria [10] or complexes which span two lipid bilayers at gap junctions,[11] and for this reason it would be desirable to be able to create double bilayer mimics as an extension of the supported bilayer field. It has been previously demonstrated that one can create such structures using the Langmuir–Blodgett technique,[12] however, this has the drawback that it is not compatible with the incorporation of transmembrane proteins and the film is created by transfer through the air–water interface. Hence approaches based on self-assembly from vesicles would provide a significant advance in the type of applications that double bilayers could be used for. Murray et al. have recently demonstrated, a previously observed phenomenon, that a second bilayer can be assembled on top of a streptavidin protein film, attached to a first bilayer.[13] Further, Chung et al. have also shown that giant unilamellar vesicle (GUV) rupture can lead to the formation of a second bilayer tethered to a first bilayer using complementary DNA sequences.[14] Similarly, GUV rupture onto a lipid bilayer to form model intermembrane junctions has been reported by Kaizuka and Groves;[15] and Tabaei et al. have demonstrated a method whereby DNA duplexes were used to tether multiple diskshaped lipid “bicelles” to a bilayer.[16] While these systems could not be used for studying double bilayer phenomena directly, they demonstrate that the principle of achieving double bilayers via self-assembly is feasible. Here we describe a new method to form double bilayer lipid membranes (dBLMs) on solid supports using NHS/EDC [17] chemistry [hydroxy-2, 5-dioxopyrrolidine-3-sulfonicacid sodium salt (NHS) and N-Ethyl-Nо-(3-dimethylaminopropyl) carbodiimide hydrochloride(EDC)