Hein Leanne K, Duplock Stephen, Hopwood John J, Fuller Maria
Lysosomal Diseases Research Unit, Department of Genetic Medicine, Children, Youth and Women's Health Service, North Adelaide, South Australia.
J Lipid Res. 2008 Aug;49(8):1725-34. doi: 10.1194/jlr.M800092-JLR200. Epub 2008 Apr 21.
The formation of cholesterol and sphingolipids into specialized liquid-ordered membrane microdomains (rafts) has been proposed to function in the intracellular sorting and transport of proteins and lipids. Defined by biochemical criteria, rafts resist solubilization in nonionic detergents, enabling them to be isolated as detergent-resistant membranes (DRM). In this study, we characterized the lipid composition of DRM from a cell model of the sphingolipid storage disorder, Gaucher disease, in which the catabolism of the sphingolipid glucosylceramide (GC) is impaired. In this cell model, we showed that GC accumulated primarily in the DRM, with smaller secondary increases in ceramide, dihexosylceramide, trihexosylceramide, and phosphatidylglycerol. This suggested that not only was lipid metabolism altered as a consequence of the cells' inability to degrade GC, but this affected the DRM rather than other regions of the membrane. This increase in lipids in the DRM may be responsible for the altered lipid and protein sorting seen in Gaucher disease. Analysis of individual lipid species revealed preservation of the shorter and fully saturated fatty acid species in the DRM, suggesting that the highly ordered and tightly packed nature of the DRM is maintained.
胆固醇和鞘脂形成特殊的液晶有序膜微区(脂筏),被认为在蛋白质和脂质的细胞内分选与运输中发挥作用。根据生化标准定义,脂筏在非离子去污剂中具有抗溶解性,这使得它们能够作为抗去污剂膜(DRM)被分离出来。在本研究中,我们对来自鞘脂贮积病戈谢病细胞模型的DRM的脂质组成进行了表征,在该模型中鞘脂葡萄糖神经酰胺(GC)的分解代谢受损。在这个细胞模型中,我们发现GC主要在DRM中积累,同时神经酰胺、二己糖神经酰胺、三己糖神经酰胺和磷脂酰甘油也有较小程度的二次增加。这表明,不仅由于细胞无法降解GC导致脂质代谢发生改变,而且这种改变影响的是DRM而非膜的其他区域。DRM中脂质的这种增加可能是戈谢病中所见脂质和蛋白质分选改变的原因。对单个脂质种类的分析显示,DRM中较短且完全饱和的脂肪酸种类得以保留,这表明DRM高度有序且紧密堆积的性质得以维持。