Hui D Y, Innerarity T L, Milne R W, Marcel Y L, Mahley R W
J Biol Chem. 1984 Dec 25;259(24):15060-8.
The ability of apolipoprotein (apo-) B48 to interact with lipoprotein receptors was investigated using three different types of lipoproteins. First, canine chylomicron remnants, which contained apo-B48 as their primary apoprotein constituent, were generated by the hydrolysis of chylomicrons with milk lipoprotein lipase. These apo-B48-containing chylomicron remnants are deficient in apo-E and reacted very poorly with apo-E receptors on adult dog liver membranes and the low density lipoprotein (apo-B,E) receptors on human fibroblasts. Addition of normal human apo-E3 restored the receptor binding activity of these lipoproteins. Second, beta-very low density lipoproteins (beta-VLDL) from cholesterol-fed dogs were subfractionated into distinct classes containing apo-E along with either apo-B48 or apo-B100. Both classes bound to the apo-B,E and apo-E receptors. Their binding was almost completely mediated by apo-E, as evidenced by the ability of the anti-apo-E to inhibit the receptor interaction. Third, beta-VLDL from type III hyperlipoproteinemic patients were subfractionated by immunoaffinity chromatography into lipoproteins containing apo-E plus either apo-B48 or apo-B100. Both subfractions bound poorly to apo-B,E and apo-E receptors due to the presence of defective apo-E2. However, the residual binding of the apo-B48-containing and apo-B100-containing human beta-VLDL was inhibited by the anti-apo-E. After lipase hydrolysis, apo-B100 became a more prominant determinant responsible for mediating receptor binding to the apo-B,E receptor. By contrast, lipase hydrolysis did not increase the binding activity of the apo-B48-containing beta-VLDL. These results indicate that apo-B48 does not play a direct role in mediating the interaction of lipoproteins with receptors on fibroblasts or liver membranes.
利用三种不同类型的脂蛋白研究了载脂蛋白(apo-)B48与脂蛋白受体相互作用的能力。首先,通过用乳脂蛋白脂肪酶水解乳糜微粒生成犬乳糜微粒残粒,其主要载脂蛋白成分是apo-B48。这些含apo-B48的乳糜微粒残粒缺乏apo-E,与成年犬肝细胞膜上的apo-E受体以及人成纤维细胞上的低密度脂蛋白(apo-B,E)受体反应很差。添加正常人apo-E3可恢复这些脂蛋白的受体结合活性。其次,将胆固醇喂养犬的β-极低密度脂蛋白(β-VLDL)分级分离成不同类别,这些类别含有apo-E以及apo-B48或apo-B100。两类都与apo-B,E和apo-E受体结合。抗apo-E抑制受体相互作用的能力证明,它们的结合几乎完全由apo-E介导。第三,通过免疫亲和色谱法将III型高脂蛋白血症患者的β-VLDL分级分离成含有apo-E加apo-B48或apo-B100的脂蛋白。由于存在缺陷的apo-E2,这两个亚组分与apo-B,E和apo-E受体的结合都很差。然而,抗apo-E抑制了含apo-B48和含apo-B100的人β-VLDL的残余结合。脂肪酶水解后,apo-B100成为介导受体与apo-B,E受体结合的更主要决定因素。相比之下,脂肪酶水解并未增加含apo-B48的β-VLDL的结合活性。这些结果表明,apo-B48在介导脂蛋白与成纤维细胞或肝细胞膜上的受体相互作用中不发挥直接作用。