title: More Than Just Oligomannose: An N-glycomic Comparison of Penicillium Species.
abstract: N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. In this study, we have compared the glycans released by peptide/N-glycosidase F from proteolysed cell pellets of three Penicillium species (P. dierckxii, P. nordicum and P. verrucosum that all belong to the Eurotiomycetes). Although the major structures are all within the range Hex(5-11)HexNAc(2) as shown by mass spectrometry, variations in reversed-phase chromatograms and MS/MS fragmentation patterns are indicative of differences in the actual structure. Hydrofluoric acid and mannosidase treatments revealed that the oligomannosidic glycans were not only in part modified with phosphoethanolamine residues and outer chain och1-dependent mannosylation, but that bisecting galactofuranose was present in a species-dependent manner. These data are the first to specifically show the modification of N-glycans in fungi with zwitterionic moieties. Furthermore, our results indicate that mere mass spectrometric screening is insufficient to reveal the subtly complex nature of N-glycosylation even within a single fungal genus.
authors.affiliation: From the ‡Department für Chemie, Universität für Bodenkultur, Muthgasse 18, 1190 Wien, Austria;
authors.forename: Alba
authors.initials: A
authors.lastname: Hykollari
authors.affiliation: From the ‡Department für Chemie, Universität für Bodenkultur, Muthgasse 18, 1190 Wien, Austria;
authors.forename: Barbara
authors.identifier: http://orcid.org/0000-0003-0818-867X
authors.initials: B
authors.lastname: Eckmair
authors.affiliation: From the ‡Department für Chemie, Universität für Bodenkultur, Muthgasse 18, 1190 Wien, Austria;
authors.forename: Josef
authors.identifier: http://orcid.org/0000-0002-4096-4926
authors.initials: J
authors.lastname: Voglmeir
authors.affiliation: §Institutionen för Biomedicin, Göteborgs universitet, 405 30 Göteborg, Sweden;
authors.forename: Chunsheng
authors.initials: C
authors.lastname: Jin
authors.affiliation: From the ‡Department für Chemie, Universität für Bodenkultur, Muthgasse 18, 1190 Wien, Austria;
authors.forename: Shi
authors.identifier: http://orcid.org/0000-0003-4949-9140
authors.initials: S
authors.lastname: Yan
authors.affiliation: From the ‡Department für Chemie, Universität für Bodenkultur, Muthgasse 18, 1190 Wien, Austria;
authors.forename: Jorick
authors.initials: J
authors.lastname: Vanbeselaere
authors.affiliation: ¶VetOMICS Core Facility, Veterinärmedizinische Universität, 1210 Wien, Austria.
authors.forename: Ebrahim
authors.initials: E
authors.lastname: Razzazi-Fazeli
authors.affiliation: From the ‡Department für Chemie, Universität für Bodenkultur, Muthgasse 18, 1190 Wien, Austria;
authors.forename: Iain B H
authors.identifier: http://orcid.org/0000-0001-8996-1518
authors.initials: IB
authors.lastname: Wilson
authors.affiliation: From the ‡Department für Chemie, Universität für Bodenkultur, Muthgasse 18, 1190 Wien, Austria; katharina.paschinger@boku.ac.at.
authors.forename: Katharina
authors.identifier: http://orcid.org/0000-0002-3594-7136
authors.initials: K
authors.lastname: Paschinger
mesh_terms: D056148:Chromatography, Reverse-Phase; D004983:Ethanolamines / Q000378:metabolism; D054794:Glycomics / Q000379:methods*; D006031:Glycosylation; D006858:Hydrofluoric Acid / Q000378:metabolism; D008358:Mannose / Q000378:metabolism*; D008361:Mannosidases / Q000378:metabolism; D009844:Oligosaccharides / Q000378:metabolism*; D010407:Penicillium / Q000145:classification / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D013045:Species Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry
chemical_list: D004983:Ethanolamines; D009844:Oligosaccharides; D011134:Polysaccharides; C005448:phosphorylethanolamine; D008361:Mannosidases; D008358:Mannose; D006858:Hydrofluoric Acid