title: Dynamic developmental elaboration of N-linked glycan complexity in the Drosophila melanogaster embryo.
abstract: The structural diversity of glycoprotein N-linked oligosaccharides is determined by the expression and regulation of glycosyltransferase activities and by the availability of the appropriate acceptor/donor substrates. Cells in different tissues and in different developmental stages utilize these control points to manifest unique glycan expression patterns in response to their surroundings. The activity of a Toll-like receptor, called Tollo/Toll-8, induces a pattern of incompletely defined, but neural specific, glycan expression in the Drosophila embryo. Understanding the full extent of the changes in glycan expression that result from altered Tollo/Toll-8 signaling requires characterization of the complete N-linked glycan profile of both wild-type and mutant embryos. N-Linked glycans harvested from wild-type or mutant embryos were subjected to direct structural analysis by analytic and preparative high pressure liquid chromatography, by multidimensional mass spectrometry, and by exoglycosidase digestion, revealing a predominance of high mannose and paucimannose glycans. Di-, mono-, and nonfucosylated forms of hybrid, complex biantennary, and triantennary glycans account for 12% of the total wild-type glycan profile. Two sialylated glycans bearing N-acetylneuraminic acid were detected, the first direct demonstration of this modification in Drosophila. Glycan profiles change during normal development consistent with increasing alpha-mannosidase II and core fucosyl-transferase enzyme activities, and with decreasing activity of the Fused lobes processing hexosaminidase. In tollo/toll-8 mutants, a dramatic, expected loss of difucosylated glycans is accompanied by unexpected decreases in monofucosylated and nonfucosylated hybrid glycans and increases in some nonfucosylated paucimannose and biantennary glycans. Therefore, tollo/toll-8 signaling influences flux through several processing steps that affect the maturation of N-linked glycans.
authors.affiliation: Complex Carbohydrate Research Center, University of Georgia, Athens, Georgia 30602, USA.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Aoki
authors.forename: Mindy
authors.initials: M
authors.lastname: Perlman
authors.forename: Jae-Min
authors.initials: JM
authors.lastname: Lim
authors.forename: Rebecca
authors.initials: R
authors.lastname: Cantu
authors.forename: Lance
authors.initials: L
authors.lastname: Wells
authors.forename: Michael
authors.initials: M
authors.lastname: Tiemeyer
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D004331:Drosophila melanogaster; D004625:Embryo, Nonmammalian; D005643:Fucose / Q000737:chemistry; D018507:Gene Expression Regulation, Developmental; D008358:Mannose / Q000737:chemistry; D013058:Mass Spectrometry; D008954:Models, Biological; D008956:Models, Chemical; D009154:Mutation; D011134:Polysaccharides / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors; D051200:Toll-Like Receptor 8 / Q000235:genetics / Q000502:physiology*
chemical_list: D011134:Polysaccharides; D051200:Toll-Like Receptor 8; D005643:Fucose; D008358:Mannose