Fishman P H
Chem Phys Lipids. 1986 Dec 15;42(1-3):137-51. doi: 10.1016/0009-3084(86)90049-6.
The recent development of several new approaches has proven extremely useful in identifying functions for gangliosides, the sialic-acid containing glycosphingolipids. The first is the incorporation of exogenous gangliosides into the plasma membrane of ganglioside-deficient cells. Using this approach, specific gangliosides have been identified as the receptors for certain bacterial toxins and viruses and as important factors in the organization of fibronectin into an extracellular matrix. The second approach has been a ligand blotting technique which allows detection of ganglioside-binding proteins such as toxins and antibodies. Gangliosides are separated by thin-layer chromatography and overlain with the protein of interest. Specific binding of the ligand to gangliosides can then be detected by either direct or indirect methods. The third approach is the use of the B or binding subunit of cholera toxin as a specific probe for endogenous plasma membrane ganglioside function. The ability of the B subunit to alter the growth of cells directly demonstrates a role for gangliosides as biotransducers of signals for the regulation of cell growth.
最近开发的几种新方法已被证明在确定神经节苷脂(含唾液酸的糖鞘脂)的功能方面极其有用。第一种方法是将外源性神经节苷脂整合到缺乏神经节苷脂的细胞的质膜中。使用这种方法,特定的神经节苷脂已被确定为某些细菌毒素和病毒的受体,以及纤连蛋白组织成细胞外基质的重要因素。第二种方法是配体印迹技术,它可以检测神经节苷脂结合蛋白,如毒素和抗体。神经节苷脂通过薄层色谱法分离,然后与感兴趣的蛋白质覆盖。然后可以通过直接或间接方法检测配体与神经节苷脂的特异性结合。第三种方法是使用霍乱毒素的B或结合亚基作为内源性质膜神经节苷脂功能的特异性探针。B亚基直接改变细胞生长的能力直接证明了神经节苷脂作为调节细胞生长信号的生物传感器的作用。