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首页> 外文期刊>Biochimica et Biophysica Acta. General Subjects >Effect of L-fucose and D-glucose concentration on L-fucoprotein metabolism in human Hep G2 cells and changes in fucosyltransferase and α-L-fucosidase activity in liver of diabetic rats
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Effect of L-fucose and D-glucose concentration on L-fucoprotein metabolism in human Hep G2 cells and changes in fucosyltransferase and α-L-fucosidase activity in liver of diabetic rats

机译：L-岩藻糖和D-葡萄糖浓度对糖尿病大鼠肝L-岩藻糖蛋白代谢及岩藻糖基转移酶和α-L-岩藻糖苷酶活性的影响

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L-Fucose is a monosaccharide that is present at low concentrations in serum and is a normal constituent of glycoproteins. In some pathological conditions, such as cancer, rheumatoid arthritis, and diabetes, there is an abnormal fucosylation of acute phase serum proteins. Because most serum proteins are produced in the liver, we have examined L-fucose accumulation, metabolism, and secretion of -fucose-containing proteins in human Hep G2 liver cells. Accumulation of L-fucose by Hep G2 cells approached 3.5 nmol/mg protein after a 48 h incubation. This accumulation appears similar to accumulation in other cells, which we have shown occurs via a specific transport protein. Exogenous -fucose was incorporated into protein in both O- and N-linked glycosidic linkages. After a 48 h incubation, 61% of the accumulated L-fucose was incorporated into protein and secreted into the medium, whereas 39% of the L-fucose remaining in the cells was incorporated into integral membrane proteins. Utilizing reverse-phase high-performance liquid chromatographic separation of L-[5,6-3H]fucose-containing proteins and detection by scintillation counting, we determined that two major fucoproteins and numerous minor fucoproteins were produced and secreted by normal Hep G2 cells. This elution profile was unchanged when glucose-conditioned cells were examined. By size-separating secreted proteins by nondenaturing HPLC we determined that the size of the two major fucoproteins were ≈60 and ≈100 kDa. In these studies we also examined the effect of diabetes on hepatic fucosyltransferase and serum α-L-fucosidase activity and found that the activity of these enzymes is increased by 40 and 100%, respectively in diabetic rats.
机译：L-岩藻糖是单糖，以低浓度存在于血清中，是糖蛋白的正常组成部分。在某些病理状况下，例如癌症，类风湿性关节炎和糖尿病，急性期血清蛋白的岩藻糖基化异常。因为大多数血清蛋白是在肝脏中产生的，所以我们检查了人类Hep G2肝细胞中L-岩藻糖的积累，代谢和含-岩藻糖的蛋白质的分泌。孵育48小时后，Hep G2细胞对L-岩藻糖的积累接近3.5 nmol / mg蛋白。这种积累似乎与其他细胞中的积累类似，我们已经证明这是通过特定的转运蛋白发生的。外源岩藻糖以O-和N-连接的糖苷键结合到蛋白质中。孵育48小时后，将61％的累积L-岩藻糖掺入蛋白质并分泌到培养基中，而将细胞中剩余的39％的L-岩藻糖掺入完整的膜蛋白中。利用反相高效液相色谱法分离含有L- [5,6-3H]岩藻糖的蛋白质并通过闪烁计数检测，我们确定正常Hep G2细胞产生并分泌了两种主要的岩藻糖蛋白和许多次要的岩藻糖蛋白。当检查葡萄糖调节的细胞时，该洗脱曲线不变。通过非变性HPLC分离分泌蛋白的大小，我们确定两种主要岩藻蛋白的大小分别为≈60和≈100kDa。在这些研究中，我们还检查了糖尿病对肝岩藻糖基转移酶和血清α-L-岩藻糖苷酶活性的影响，发现这些酶的活性在糖尿病大鼠中分别增加了40％和100％。

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