title: Glycosylation signatures in Drosophila: fishing with lectins.
abstract: Glycosylation is a co- and/or post-translational protein modification that generates enormous structural diversity among glycoproteins. In this study, immobilized lectins were used to capture glycoproteins with different glycan profiles from Drosophila melanogaster extracts. On the basis of previous results from glycan array analyses, the snowdrop (Galanthus nivalis) agglutinin (GNA), the tobacco (Nicotiana tabacum) lectin (Nictaba) and the Rhizoctoni solani agglutinin (RSA) were used to select for a broad range of N- and O-glycan structures. After different lectin affinity chromatographies, the glycoproteome of Drosophila was analyzed using LC-MS/MS and glycoprotein abundances were calculated by different label-free methods. Bioinformatics tools were used to annotate the identified glycoproteins and the glycoproteins were classified according to their molecular function or their involvement in a biological process. Subsequent enrichment analysis (using the DAVID database) was employed to find biological processes or molecular functions in Drosophila in which a particular glycan signature is overrepresented. The results presented here clearly demonstrate that next to the presence of high-mannose and pauci-mannose N-glycans, Drosophila is capable of synthesizing glycoproteins carrying extended hybrid and complex N-linked glycans. Furthermore, it was demonstrated that a specific glycosylation signature can be associated with a functionally related group of glycoproteins in Drosophila, both in terms of biological process and molecular function.
authors.affiliation: Laboratory of Agrozoology, Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, Coupure links 653, Ghent, Belgium.
authors.forename: Gianni
authors.initials: G
authors.lastname: Vandenborre
authors.forename: Els J M
authors.initials: EJ
authors.lastname: Van Damme
authors.forename: Bart
authors.initials: B
authors.lastname: Ghesquière
authors.forename: Gerben
authors.initials: G
authors.lastname: Menschaert
authors.forename: Mohamad
authors.initials: M
authors.lastname: Hamshou
authors.forename: Rameshwaram Nagender
authors.initials: RN
authors.lastname: Rao
authors.forename: Kris
authors.initials: K
authors.lastname: Gevaert
authors.forename: Guy
authors.initials: G
authors.lastname: Smagghe
mesh_terms: D000818:Animals; D019295:Computational Biology / Q000379:methods; D029721:Drosophila Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000737:chemistry / Q000378:metabolism*; D006023:Glycoproteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006031:Glycosylation; D055767:Immobilized Proteins / Q000378:metabolism*; D037121:Plant Lectins / Q000378:metabolism*
chemical_list: D029721:Drosophila Proteins; D006023:Glycoproteins; D055767:Immobilized Proteins; D037121:Plant Lectins