Identification of N-Glycosylated Proteins from the Central Nervous System of Drosophila Melanogaster

作者：Kate Koles, Jong‐Min Lim, Kazuhiro Aoki, M. Porterfield, Michael Tiemeyer, Lance Wells, Vladsilav M Panin · 发表于：Glycobiology · 年份：2007 · DOI：10.1093/glycob/cwm097 · 被引用次数：101 · 研究领域：Glycosylation and Glycoproteins Research、Invertebrate Immune Response Mechanisms、Studies on Chitinases and Chitosanases

Although the function of many glycoproteins in the nervous system of fruit flies is well understood, information about the glycosylation profile and glycan attachment sites for such proteins is scarce. In order to fill this gap and to facilitate the analysis of N-linked glycosylation in the nervous system, we have performed an extensive survey of membrane-associated glycoproteins and their N-glycosylation sites isolated from the adult Drosophila brain. Following subcellular fractionation and trypsin digestion, we used different lectin affinity chromatography steps to isolate N-glycosylated glycopeptides. We identified a total of 205 glycoproteins carrying N-linked glycans and revealed their 307 N-glycan attachment sites. The size of the resulting dataset furthermore allowed the statistical characterization of amino acid distribution around the N-linked glycosylation sites. Glycan profiles were analyzed separately for glycopeptides that were strongly and weakly bound to Concanavalin A (Con A), or that failed to bind Concanavalin A, but did bind to wheat germ agglutinin (WGA). High- or paucimannosidic glycans dominated each of the profiles, although the wheat germ agglutinin-bound glycan population was enriched in more extensively processed structures. A sialylated glycan structure was unambiguously detected in the wheat germ agglutinin-bound fraction. Despite the large amount of starting material, insufficient amount of glycopeptides was retained by the Wisteria floribunda (WF...