Variations in glycosylation of von Willebrand factor with O-linked sialylated T antigen are associated with its plasma levels

Blood. 2007 Mar 15;109(6):2430-7. doi: 10.1182/blood-2006-06-032706. Epub 2006 Nov 7.

Abstract

The glycosylation profile of von Willebrand factor (VWF) is known to strongly influence its plasma levels. VWF contains several carbohydrate structures, including O-linked glycans that primarily consist of sialylated T antigen (NeuAc(alpha2-3)Gal-(beta1-3)-[NeuAc(alpha2-6)]GalNAc). It is not yet known whether O-linked carbohydrates affect VWF levels. We developed an immunosorbent assay based on neuraminidase incubation allowing subsequent binding of peanut agglutinin (PNA) to desialylated O-linked T antigen on VWF. An inverse relation was found between PNA binding and VWF antigen levels in healthy individuals (n = 111; Pearson rank = -0.43; P < .001). A similar inverse association was observed in randomly selected plasma samples from our diagnostic laboratory: 252% +/- 125% for VWF levels less than 0.5 U/mL (n = 15); 131% +/- 36% for VWF levels between 0.5 and 1.5 U/mL (n = 32); and 92% +/- 40% for VWF levels more than 1.5 U/mL (n = 19). Reduced or increased PNA binding was also observed in patients with increased (liver cirrhosis) or reduced (von Willebrand disease [VWD] type 1) VWF antigen levels, respectively. VWD type 1 patients further displayed increased ratios of propeptide over mature VWF antigen levels (0.38 +/- 0.18 versus 0.17 +/- 0.03 for patients and controls, respectively; P < .001), which is indicative of reduced VWF survival in these patients. Of interest, a linear relation between PNA binding and propeptide/VWF ratio was observed (Spearman rank = 0.47), suggesting a potential association between O-linked glycosylation and VWF survival. Finally, we detected a marked decrease in PNA binding in post-DDAVP (1-deamino-8-D-arginine vasopressin) samples from various patients, indicating that the O-linked glycosylation profile of VWF stored in endothelial storage organelles may differ from circulating VWF.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Viral, Tumor / blood*
  • Antigens, Viral, Tumor / genetics*
  • Genetic Variation / genetics*
  • Glycosylation
  • Humans
  • Liver Cirrhosis / blood
  • Liver Cirrhosis / genetics
  • N-Acetylneuraminic Acid / metabolism*
  • Peanut Agglutinin / metabolism
  • Protein Binding
  • Protein Processing, Post-Translational
  • von Willebrand Factor / chemistry
  • von Willebrand Factor / genetics*
  • von Willebrand Factor / metabolism*

Substances

  • Antigens, Viral, Tumor
  • Peanut Agglutinin
  • von Willebrand Factor
  • N-Acetylneuraminic Acid