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

• DOI: 10.1074/jbc.m804925200
• Publication Date: 2008-08-26T00:45:03Z
• Authors: Kazuhiro Aoki; Mindy Porterfield; Samuel S. Lee; Brian Dong; Khoi Nguyen; Katherine H. McGl...; Michael Tiemeyer
• Keywords: Trisaccharide; Glycomics; Exoglycosidase; Lumican; Disaccharide

Appropriate glycoprotein O-glycosylation is essential for normal development and tissue function in multicellular organisms. To comprehensively assess the developmental functional impact of altered O-glycosylation, we have extensively analyzed non-glycosaminoglycan, O-linked glycans expressed Drosophila embryos. Through multidimensional mass spectrometric analysis released from glycoproteins by beta-elimination, detected novel as well previously reported O-glycans that exhibit developmentally modulated expression. The core 1 mucin-type disaccharide (Galbeta1-3GalNAc) predominant glycan total profile. HexNAcitol, hexitol, xylosylated branching extension with HexNAc (to generate 2 glycans) were also evident following release reduction. After Galbeta1-3GalNAc, next most prevalent a mixture novel, isobaric, linear, branched forms glucuronyl disaccharide. Other less structures extended HexA, including an O-fucose glycan. Although expected product Fringe glycosyltransferase, (GlcNAcbeta1-3)fucitol, was not detectable whole embryos, spectrometry fragmentation exoglycosidase sensitivity defined trisaccharide GlcNAcbeta1-3(GlcAbeta1-4)fucitol. Consistent spatial distribution function, GlcA-extended form enriched dorsal portion wing imaginal disc. Furthermore, loss activity reduced prevalence O-Fuc trisaccharide. Therefore, necessary modulation important signaling events are, vertebrates, substrates beyond addition single HexNAc.