Schneider W J
J Cell Biochem. 1983;23(1-4):95-106. doi: 10.1002/jcb.240230110.
The receptor for low density lipoprotein was purified from bovine adrenal cortex in the presence of the nonionic detergent octylglucoside. Receptors were incorporated into the bilayer of egg phosphatidylcholine vesicles by a detergent-dialysis method. Reconstituted receptors were functional in that they bound low density lipoprotein as well as a monoclonal antibody directed against the receptor in a specific, saturable fashion. Binding activity of reconstituted receptors was measured by a gel chromatography assay. The orientation of the receptor molecule within the phospholipid bilayer was investigated by binding assays following proteolytic digestion. Reconstituted receptors showed an orientation that was functionally indistinguishable from that of low density lipoprotein receptors in the plasma membrane of intact human fibroblasts.
低密度脂蛋白受体是在非离子去污剂辛基葡糖苷存在的情况下从牛肾上腺皮质中纯化出来的。通过去污剂透析法将受体整合到卵磷脂酰胆碱囊泡的双层膜中。重构后的受体具有功能，因为它们能以特异性、可饱和的方式结合低密度脂蛋白以及针对该受体的单克隆抗体。通过凝胶色谱分析来测定重构受体的结合活性。通过蛋白水解消化后的结合试验研究了受体分子在磷脂双层膜中的取向。重构后的受体显示出一种取向，在功能上与完整人成纤维细胞质膜中的低密度脂蛋白受体的取向没有区别。
