Two-dimensional gel electrophoresis of apolipoprotein C-III and other serum glycoproteins for the combined screening of human congenital disorders of a and N-glycosylation
Two-dimensional gel electrophoresis of apolipoprotein C-III and other serum glycoproteins for the combined screening of human congenital disorders of a and N-glycosylation
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DOI：
10.1002/prca.200600777
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发表时间：
2007-03-01
影响因子：
2
通讯作者：
Seta, Nathalie
中科院分区：
文献类型：
--
作者：
Bruneel, Arnaud;Robert, Tiphaine;Seta, Nathalie
先天性糖基化障碍（CDG）是一种遗传性疾病，不仅影响N-聚糖（如CDG I型和II型），还影响O-聚糖生物合成途径。在没有特定临床症状的情况下，需要对这两组CDG进行可靠的生物学筛查。使用几微升的人血清，2-DE和免疫印迹应用于分离和同时检测的O-糖基化的蛋白质载脂蛋白C-III（apoC-III）和四个N-糖基化的蛋白质，即α-抗胰蛋白酶，α-1酸性糖蛋白，触珠蛋白和转铁蛋白的亚型。对于O-糖基化的研究，该技术允许可靠的分离apoC-III的三个组分，并确定成人人群中的正常百分比值。关于N-糖基化，对Ia型CDG患者血清样本的研究显示，系统性影响四种2-DE分离的N-连接糖蛋白的显着异常。2-DE偶联免疫印迹使用的特异性抗体的混合物可以很容易地和可靠地用于在人类的N-和O-糖基化疾病的联合筛查。
Congenital disorders of glycosylation (CDG) are inherited diseases that can affect not only the N-glycan (e.g. CDG type I and II) but also the O-glycan biosynthesis pathway. In the absence of specific clinical symptoms, there is a need for a reliable biological screening of these two groups of CDG. Using a few microlitres of human serum, 2-DE and immunoblotting were applied to the separation and simultaneous detection of the isoforms of the O-glycosylated protein apolipoprotein C-III (apoC-III) and of four N-glycosylated proteins, namely alpha-antitrypsin, alpha-1 acid glycoprotein, haptoglobin and transferrin. For the study of O-glycosylation, this technique allowed the reliable separation of the three fractions of apoC-III and the determination of normal percentage values in an adult population. Concerning N-glycosylation, the study of serum samples from patients with CDG type Ia revealed marked abnormalities systematically affecting the four 2-DE separated N-linked glycoproteins. 2-DE coupled to immunoblotting using a mixture of specific antibodies could be easily and reliably employed for the combined screening of both N- and O-glycosylation disorders in humans.