Abstract
Antennary fucosylation alterations in plasma glycoproteins have been previously proposed and tested as a biomarker for differentiation of maturity onset diabetes of the young (MODY) patients carrying a functional mutation in the HNF1A gene. Here, we developed a novel LC-based workflow to analyze blood plasma N-glycan fucosylation in 320 diabetes cases with clinical features matching those at risk of HNF1A-MODY. Fucosylation levels measured in two independent research centers by using similar LC-based methods were correlated to evaluate the interlaboratory performance of the biomarker. The interlaboratory study showed good correlation between fucosylation levels measured for the 320 cases in the two centers with the correlation coefficient (r) of up to 0.88 for a single trait A3FG3S2. The improved chromatographic separation allowed the identification of six single glycan traits and a derived antennary fucosylation trait that were able to differentiate individuals carrying pathogenic mutations from benign or no HNF1A mutation cases, as determined by the area under the curve (AUC) of up to 0.94. The excellent (r = 0.88) interlaboratory performance of the glycan biomarker for HNF1A-MODY further supports the development of a clinically relevant diagnostic test measuring antennary fucosylation levels.
Highlights
Glycosylation is a co-/post-translational modification which affects protein conformation and interactions
The research presented here was performed using blood plasma samples obtained from study participants that were recruited via the Young Diabetes in Oxford (YDX) study in the UK (n = 90) and the Croatian National Diabetes Registry (CroDiab) in Croatia (n = 230)
To determine the antennary fucosylation levels of 320 subjects with diabetes, we developed a workflow encompassing highly reproducible ultra-high-performance liquid chromatography (UHPLC) analysis and high-throughput (HTP) systematic data processing
Summary
Glycosylation is a co-/post-translational modification which affects protein conformation and interactions. Glycans can be covalently attached to proteins through either N- or Oglycosidic linkages. These oligosaccharide chains strongly influence protein-protein interactions and are involved in protein folding, sub-cellular targeting, and trafficking. N-glycans are linked to the amide group of asparagine (Asn), whereas Oglycans are attached to the hydroxyl group of serine (Ser) or threonine (Thr) in the polypeptide backbone [1]. Aberrant protein glycosylation has been associated with many pathological conditions in humans. Some chronic inflammatory [2], autoimmune [3] and infectious diseases [4], Center, Leiden, The Netherlands 3 Quadram Institute Bioscience, Norwich Research Park, Norwich, UK 4 Genos Glycoscience Research Laboratory, Zagreb, Croatia
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