A rapid mass spectrometric strategy for the characterization of N- and O-glycan chains in the diagnosis of defects in glycan biosynthesis
A rapid mass spectrometric strategy for the characterization of N- and O-glycan chains in the diagnosis of defects in glycan biosynthesis
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DOI：
10.1002/pmic.200600977
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发表时间：
2007-06-01
期刊：
影响因子：
3.4
通讯作者：
Morelle, Willy
中科院分区：
文献类型：
--
作者：
Faid, Valegh;Chirat, Frederic;Morelle, Willy
蛋白质的糖基化是一个非常复杂的过程，涉及许多因素，如酶或转运蛋白。聚糖生物合成途径中这些因子之一的缺陷导致称为先天性糖基化障碍（CDG）的严重疾病。CDG不仅可以影响蛋白质N-聚糖的生物合成，还可以影响O-聚糖的生物合成。血清糖蛋白聚糖的结构分析是解决这一缺陷的关键。出于这个原因，我们在本文中提出了一个战略，同时表征的N-和O-聚糖链分离的血清糖蛋白。血清（20 μ L）用于表征通过PNGase F酶消化释放的N-聚糖。使用10 μ L血清，通过还原消除从全血清糖蛋白中化学释放O-聚糖。使用基于质谱分析的策略，定义N-和O-聚糖链的结构。这些策略应用于一名患者的血清与CDG IIa型，和一名患者的轻度形式的先天性糖基化II型（CDG-II），这是由Cog 1亚基的复合物的缺陷引起的。
Glycosylation of proteins is a very complex process which involves numerous factors such as enzymes or transporters. A defect in one of these factors in glycan biosynthetic pathways leads to dramatic disorders named congenital disorders of glycosylation (CDG). CDG can affect the biosynthesis of not only protein N-glycans but also O-glycans. The structural analysis of glycans on serum glycoproteins is essential to solving the defect. For this reason, we propose in this paper a strategy for the simultaneous characterization of both N- and O-glycan chains isolated from the serum glycoproteins. The serum (20 mu L) is used for the characterization of N-glycans which are released by enzymatic digestion with PNGase F. O-glycans are chemically released by reductive elimination from whole serum glycoproteins using 10 mu L of the serum. Using strategies based on mass spectrometric analysis, the structures of N- and O-glycan chains are defined. These strategies were applied on the sera from one patient with CDG type IIa, and one patient with a mild form of congenital disorder of glycosylation type II (CDG-II) that is caused by a deficiency in the Cog1 subunit of the complex.