Two‐dimensional gel electrophoresis of apolipoprotein C‐III and other serum glycoproteins for the combined screening of human congenital disorders of O‐ and N‐glycosylation |
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Authors: | Arnaud Bruneel Dr Tiphaine Robert Dirk J Lefeber Guillaume Benard Emilie Loncle Amel Djedour Geneviève Durand Nathalie Seta |
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Affiliation: | 1. Assistance Publique – H?pital de Paris, Service de Biochimie Métabolique et Cellulaire, Groupe hospitalier Bichat – Claude Bernard, Paris, FranceAP‐HP, Service de Biochimie Métabolique, H?pital Bichat, 46 rue Henri Huchard, 75018 Paris, France Fax: +33‐1‐40‐25‐88‐21;2. Assistance Publique – H?pital de Paris, Service de Biochimie Métabolique et Cellulaire, Groupe hospitalier Bichat – Claude Bernard, Paris, France;3. Laboratory of Pediatrics and Neurology, Radboud University Nijmegen Medical Center, Nijmegen, The Netherlands |
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Abstract: | 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. |
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Keywords: | Apolipoprotein C‐III CDG Glycoproteins O‐glycosylation Two‐dimensional gel electrophoresis |
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