Carbohydrate analysis of human von Willebrand factor with horseradish peroxidase-conjugated lectins.
Human von Willebrand factor (vWF) immobilized on a polyvinylidene difluoride membrane was subjected to binding assay with a series of horseradish peroxidase-conjugated lectins. The protein was reactive with concanavalin A, Ricinus communis agglutinin 120, wheat germ agglutinin and Ulex europaeus agglutinin I (UEA-I) but not with peanut agglutinin before sialidase treatment. These reactivities were consistent with the major oligosaccharide structure reported except for UEA-I. The reactivity with UEA-I was greatly decreased after digestion of the protein with either alpha-L-fucosidase or peptide-N-glycosidase F, but no significant decrease was observed after mild alkaline treatment or delipidation. vWF and UEA-I have been independently used as a good marker for human endothelial cells. Our results indicate that vWF itself contains UEA-I reactive sugar chains in its Asn-linked oligosaccharides.
Matsui
T
T
Division of Biomedical Polymer Science, Fujita Health University School of Medicine, Aichi, Japan.
Kihara
C
C
Fujimura
Y
Y
Mizuochi
T
T
Titani
K
K
D002241:Carbohydrates / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000097:blood; D006026:Glycoside Hydrolases; D006735:Horseradish Peroxidase; D006801:Humans; D037102:Lectins; D008970:Molecular Weight; D009844:Oligosaccharides / Q000737:chemistry / Q000302:isolation & purification; D011485:Protein Binding; D014841:von Willebrand Factor / Q000737:chemistry* / Q000302:isolation & purification
D002241:Carbohydrates; D006023:Glycoproteins; D037102:Lectins; D009844:Oligosaccharides; D014841:von Willebrand Factor; D006735:Horseradish Peroxidase; D006026:Glycoside Hydrolases