生物物理-生物膜的离体研究.pptVIP

The formation of BLM by apposing two monolayers on the tip of a micropipette by first removing the tip from the aqueous phase on which a monolayer is present, and then reinserting the tip through the monolayer. 不对称BLM的制备(2) 在滤纸的微孔中形成BLM 在显微镜下观察滤纸，可以看到上面有许多微孔。有人把滤纸先用脂浸泡，然后取出晾干，这样滤纸就具有亲脂疏水特性。用这种脂处理过的滤纸放在聚四氟乙烯隔板的部位取代聚四氟乙烯隔板（当然要有特别设计的夹具，把滤纸夹在左右二室之间）。然后用前面介绍的方法铺制BLM，实践证明在滤纸的许多小孔上都形成了BLM，只是每个BLM的面积小，但是许多小面积的BLM加起来就比单个“大”孔上铺成的BLM还大。 生物膜的离体研究 脂单层方法 支撑脂双层方法 平面脂双层(黑脂膜)方法 脂质体方法 Temperature induced shape changes of vesicles (a) Transition of a vesicle from a biconcave shape (center) to an invaginated (stomatocyte) shape (left side) at decreasing temperature and to a budded shape (right side) at increasing temperature. Shapes of vesicles (or cells) on the basis of the bilayer coupling hypothesis model (1983): The different cell shapes can be explained in terms of two parameters: 1) the reduced volume v=v/vs: the ratio of the actual volume of the cell to that of a sphere, 2) the relative area difference ?=(A0-Ai)/(A0,s-Ai,s) where A0 and Ai are the areas of the outer and inner leaflet of the membrane and where A0,s and Ai,s are the corresponding values of the sphere. Shape transformation of giant vesicle triggered by temperature changes. The transitions are completely reversible ! Phase contrast micrographs of shape change of composite shell (1998). DMPC vesicle containing 2?M of actin (image a and b) and 7 ? M(image c and d) of actin. 脂质体的类型： SUV： 小单片层脂质体(small unilaminar-) MLV：多片层脂质体(multilayer-) LUV: 大单片层脂质体(large unilaminar-) 脂质体是由脂质双分子层组成的内部为水相的闭合囊泡。 SUV LUV MUV MVL 20-100nm 100-500nm 0.1-5?m 1-100??m 不同类型脂质体的比较 分类： 按结构 Three methods for reconstitution. (1) Lipid and proteins solubilized in detergents are mixed and then the detergent is removed by dialysis, dilution, or gel filtration. (2) Delipidated proteins can sometimes be spontaneously incorporated into