title: Profiling N-glycans of the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectrometry systems.
abstract: Sea urchin eggs are surrounded by a carbohydrate-rich layer, termed the jelly coat, that consists of polysaccharides and glycoproteins. In the present study, we describe two mass spectrometric strategies to characterize the N-glycosylation of the Paracentrotus lividus egg jelly coat, which has an alecithal-type extracellular matrix like mammalian eggs. Egg jelly was isolated, lyophilized, and dialyzed, followed by peptide N-glycosidase F (PNGase-F) treatment to release N-glycans from their protein chain. These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). N-glycans in the egg jelly coat glycoproteins were indicated by sodiated molecules at m/z 1579.8, 1783.9, 1988.0, 2192.0, and 2397.1 for permethylated oligosaccharides on MALDI-TOF MS. Fragmentation and structural characterization of these oligosaccharides were performed by ESI-Ion trap MS/MS. Then, MALDI-TOF-MS and ESI-Ion trap-MS/MS spectra were interpreted using the GlycoWorkbench software suite, a tool for building, displaying, and profiling glycan masses, to identify the original oligosaccharide structures. The oligosaccharides of the isolated egg jelly coat were mainly of the high mannose type.
authors.affiliation: Faculty of Science, Department of Biology, Molecular Biology Section, Ege University, Bornova, İzmir.
authors.forename: Umut
authors.identifier: 0000-0002-2200-6986
authors.initials: U
authors.lastname: Şahar
authors.affiliation: Faculty of Science, Department of Biology, Molecular Biology Section, Ege University, Bornova, İzmir.
authors.forename: Remziye
authors.initials: R
authors.lastname: Deveci
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid; D005109:Extracellular Matrix / Q000737:chemistry / Q000378:metabolism*; D005260:Female; D010063:Ovum / Q000737:chemistry / Q000378:metabolism*; D047328:Paracentrotus / Q000737:chemistry / Q000378:metabolism*; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D011134:Polysaccharides
keywords: LC-MS/MS; N-glycosylation; egg jelly; fertilization; glycomics