HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 283, Issue 44, 30385-30400, October 31, 2008

This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: 283/44/30385    most recent M804925200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Aoki, K. Articles by Tiemeyer, M. Search for Related Content PubMed PubMed Citation Articles by Aoki, K. Articles by Tiemeyer, M. Social Bookmarking                      What's this?

The Diversity of O-Linked Glycans Expressed during Drosophila melanogaster Development Reflects Stage- and Tissue-specific Requirements for Cell Signaling^*

From the Complex Carbohydrate Research Center and Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia 30602

Appropriate glycoprotein O-glycosylation is essential for normal development and tissue function in multicellular organisms. To comprehensively assess the developmental and functional impact of altered O-glycosylation, we have extensively analyzed the non-glycosaminoglycan, O-linked glycans expressed in Drosophila embryos. Through multidimensional mass spectrometric analysis of glycans released from glycoproteins by β-elimination, we detected novel as well as previously reported O-glycans that exhibit developmentally modulated expression. The core 1 mucin-type disaccharide (Galβ1-3GalNAc) is the predominant glycan in the total profile. HexNAcitol, hexitol, xylosylated hexitol, and branching extension of core 1 with HexNAc (to generate core 2 glycans) were also evident following release and reduction. After Galβ1-3GalNAc, the next most prevalent glycans were a mixture of novel, isobaric, linear, and branched forms of a glucuronyl core 1 disaccharide. Other less prevalent structures were also extended with HexA, including an O-fucose glycan. Although the expected disaccharide product of the Fringe glycosyltransferase, (GlcNAcβ1-3)fucitol, was not detectable in whole embryos, mass spectrometry fragmentation and exoglycosidase sensitivity defined a novel glucuronyl trisaccharide as GlcNAcβ1-3(GlcAβ1-4)fucitol. Consistent with the spatial distribution of the Fringe function, the GlcA-extended form of the Fringe product was enriched in the dorsal portion of the wing imaginal disc. Furthermore, loss of Fringe activity reduced the prevalence of the O-Fuc trisaccharide. Therefore, O-Fuc glycans necessary for the modulation of important signaling events in Drosophila are, as in vertebrates, substrates for extension beyond the addition of a single HexNAc.

Received for publication, June 27, 2008 , and in revised form, August 8, 2008.

^* This work was supported, in whole or in part, by National Institutes of Health Grants 1-R01-GM072839 from NIGMS and 1-U01-CA128454 from NCI (both to M. T.). This work was also supported by a Toyobo Biotechnology Foundation long-term research grant (to K. A.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. 1-8 and Tables 1-3.

¹ To whom correspondence should be addressed: The Complex Carbohydrate Research Center, University of Georgia, 315 Riverbend Rd., Athens, GA 30602-4712. Tel.: 706-542-2740; Fax: 706-542-4412; E-mail: mtiemeyer{at}ccrc.uga.edu.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				K. B. Luther, H. Schindelin, and R. S. Haltiwanger Structural and Mechanistic Insights into Lunatic Fringe from a Kinetic Analysis of Enzyme Mutants J. Biol. Chem., January 30, 2009; 284(5): 3294 - 3305. [Abstract] [Full Text] [PDF]