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Title: A fucose-containing O-glycoepitope on bovine and human nucleolin
Authors: Aldi, Silvia
Focarelli, Riccardo
Armini, Alessandro 
Ziche, Marina
Finetti, Federica 
Rosati, Floriana
Keywords: A431; human cancer cells; CVEC; glycoepitopes; nucleolin; RNA-interference
Issue Date: 2009
Project: None 
In this paper, we demonstrate the existence and localizationof fucosyl-containing O-glycoforms of nucleolin in culturedbovine endothelial cells (CVEC) and malignant culturedhuman A431 cells. The tool for this discovery was anantibody raised against gp273, a glycoprotein ligand for thesperm–egg interaction in the mollusc bivalve Unio elongatulus.The function and immunological properties of gp273mainly depend on clustered Lewis-like, fucose-containingO-glycans. Here an anti-gp273 antibody was used to evaluatewhether glycoepitopes similar to those of gp273 are partof potential ligands of selectins in endothelial cells.We foundthat anti-gp273 strongly and exclusively interacted with a110 kDa protein in CVEC and A431 tumor cells. After partialpurification, mass spectrometry identified the proteinas nucleolin. This was confirmed by comparing anti-gp273and anti-nucleolin antibody immunoblotting after nucleolindepletion.We confirmed that anti-gp273 binding to nuclearand extranuclear nucleolin was against a fucose-containingO-glycoepitope by immunoblot analysis of the protein afterchemically removing O-glycans and by lectin-blot analysisof control and nucleolin-depleted samples. Using anti-gp273IgG, we detected nucleolin on the plasma membrane andcytoplasm. O-glycosylation may regulate the plethora offunctions in which nucleolin is involved.
ISSN: 0959-6658
DOI: 10.1093/glycob/cwn126
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