Site-specific analysis of von Willebrand factor O-glycosylation

Journal of Thrombosis and Haemostasis : JTH
B A SoleckaC Kannicht

Abstract

O-glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O-glycosylation sites. The aim of this study was the site-specific analysis of VWF O-glycosylation. Tryptic VWF-O-glycopeptides were isolated by lectin affinity chromatography and/or by reverse-phase high-performance liquid chromatography. Subsequently, the purified glycopeptides were analyzed by glycosidase digestion and mass spectrometry. We found that all 10 predicted O-glycosylation sites in VWF are occupied. The majority of the glycan structures on all glycosylation sites is represented by disialyl core 1 O-glycan. The presence of core 2 O-glycan was also confirmed; interestingly, this structure was not evenly distributed among all 10 glycosylation sites. Analysis of the glycopeptides flanking the A1 domain revealed that generally more core-2-type O-glycan was present on the C-terminal Cluster 2 glycopeptide (encompassing T(1468) , T(1477) , S(1486) and T(1487) ) compared with the N-terminal Cluster 1 glycopeptide (encompassing T(1248) , T(1255) , T(1256) and S(1263) ). Disialosyl motifs were present on both glycopeptides flanking the A1 domain and on the glycosylation...Continue Reading

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Citations

Jul 12, 2017·Journal of Proteome Research·Ebtesam A GashashPeng George Wang
Nov 23, 2017·Molecular & Cellular Proteomics : MCP·Zsuzsanna Darula, Katalin F Medzihradszky
Jul 12, 2017·Journal of Thrombosis and Haemostasis : JTH·W DengR Li
Sep 4, 2019·Journal of Thrombosis and Haemostasis : JTH·Alexander TischerMatthew Auton
May 6, 2019·Journal of Thrombosis and Haemostasis : JTH·Soracha WardJames S O'Donnell
Feb 1, 2017·Hämostaseologie·Maria A Brehm
Oct 29, 2020·Scientific Reports·Diarmaid O'DonghaileJames S O'Donnell
Jun 16, 2021·Seminars in Thrombosis and Hemostasis·Soracha WardJames S O'Donnell
Sep 17, 2021·Journal of Thrombosis and Haemostasis : JTH·Kayleigh M VoosRenhao Li

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