Skip Navigation



Glycobiology Advance Access published online on October 26, 2006
Glycobiology, doi:10.1093/glycob/cwl063
This Article
Right arrow Advance Access manuscript (PDF) Freely available
Right arrow Supplementary Data
Right arrow All Versions of this Article:
17/2/141    most recent
cwl063v1
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 Sandra, K.
Right arrow Articles by Van Beeumen, J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Sandra, K.
Right arrow Articles by Van Beeumen, J.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2006. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org
Received April 27, 2006
Revised October 16, 2006
Accepted October 20, 2006

Article

New insights in Rapana venosahemocyanin N-glycosylation resulting from on-line mass spectrometric analyses

Koen Sandra 1, Pavlina Dolashka-Angelova 2, Bart Devreese 1, and Jozef Van Beeumen 3 *

1 Laboratory of Protein Biochemistry and Protein Engineering, Ghent University, 9000 Ghent, Belgium
2 Institute of Organic Chemistry, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria
3 Laboratory of Protein Biochemistry and Protein Engineering, Ghent University, K.L. Ledeganckstraat 35, B-9000 Ghent, Belgium

* To whom correspondence should be addressed.
Jozef Van Beeumen, E-mail: Jozef.vanbeeumen{at}UGent.be


  Abstract

The N-glycosylation of structural unit 1 of Rapana venosa hemocyanin (RvH1) was studied. Enzymatically liberated N-glycans were analysed by MALDI-TOF-MS and CE-MS following 8-aminopyrene-1,3,6-trisulphonate labelling and labelling with 3-aminopyrazole, a new dedicated sugar reagent. Structural information was obtained by exoglycosidase sequencing, on-line MS/MS, permethylation and amidation. A mixture of high-mannose and complex glycans with so far unknown and unusual acidic terminal structures was revealed. Since the hemocyanin protein sequence is currently unknown, de novo sequencing of the glycopeptides had to be carried out. The N-glycans were therefore enzymatically removed with simultaneous partial (50%) 18O-labelling of glycosylated asparagine residues prior to proteolysis. Following nano-LC-MALDI-TOF-MS, the originally glycosylated peptides could be revealed and their sequences determined by MS/MS. The site-occupancies were subsequently elucidated by precursor ion scanning of the intact glycopeptides using a Q-Trap mass spectrometer.

Keywords: Capillary electrophoresis-mass spectrometry (CE-MS); Glycosylation; hemocyanin; MALDI-TOF; TOF; Rapana venosa.
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.