Skip Navigation



Glycobiology Advance Access published online on November 20, 2008
Glycobiology, doi:10.1093/glycob/cwn126
This Article
Right arrow Advance Access manuscript (PDF) Freely available
Right arrow All Versions of this Article:
19/4/337    most recent
cwn126v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Disclaimer
Google Scholar
Right arrow Articles by Aldi, S.
Right arrow Articles by Rosati, F.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Aldi, S.
Right arrow Articles by Rosati, F.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2008. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

Communications

A fucose-containing O-glycoepitope on bovine and human nucleolin

Silvia Aldi1, Cinzia Della Giovampaola1, Riccardo Focarelli1, Alessandro Armini2, Marina Ziche2, Federica Finetti2 and Floriana Rosati1

1 Department of Evolutionary Biology
2 Department of Molecular Biology

All correspondence to: Prof. Floriana Rosati, Dept. Evolutionary Biology, University of Siena, Via A. Moro 2, 53100 Siena, Italy Tel: +39-0577-234408; Fax: +39-0577-234476; E-mail: rosatif{at}unisi.it

Received on December 23, 2007; accepted on November 3, 2008

In this paper, we demonstrate the existence and localization of fucosyl-containing O-glycoforms of nucleolin in cultured bovine endothelial cells (CVEC) and malignant cultured human A431 cells. The tool for this discovery was an antibody raised against gp273, a glycoprotein ligand for the sperm–egg interaction in the mollusc bivalve Unio elongatulus. The function and immunological properties of gp273 mainly depend on clustered Lewis-like, fucose-containing O-glycans. Here an anti-gp273 antibody was used to evaluate whether glycoepitopes similar to those of gp273 are part of potential ligands of selectins in endothelial cells. We found that anti-gp273 strongly and exclusively interacted with a 110 kDa protein in CVEC and A431tumor cells. After partial purification, mass spectrometry identified the protein as nucleolin. This was confirmed by comparing anti-gp273 and anti-nucleolin antibody immunoblotting after nucleolin depletion. We confirmed that anti-gp273 binding to nuclear and extranuclear nucleolin was against a fucose-containing O-glycoepitope by immunoblot analysis of the protein after chemically removing O-glycans and by lectin-blot analysis of control and nucleolin-depleted samples. Using anti-gp273 IgG, we detected nucleolin on the plasma membrane and cytoplasm. O-Glycosylation may regulate the plethora of functions in which nucleolin is involved.

Key words: nucleolin / glycoforms / glycoepitopes / O-glycans / CVEC / A431 human cancer cells / RNA-interference


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.