RESUMO
The glycosylation abilities of snails deserve attention, because snail species serve as intermediate hosts in the developmental cycles of some human and cattle parasites. In analogy to many other host-pathogen relations, the glycosylation of snail proteins may likewise contribute to these host-parasite interactions. Here we present an overview on the O-glycan structures of 8 different snails (land and water snails, with or without shell): Arion lusitanicus, Achatina fulica, Biomphalaria glabrata, Cepaea hortensis, Clea helena, Helix pomatia, Limax maximus and Planorbarius corneus. The O-glycans were released from the purified snail proteins by ß-elimination. Further analysis was carried out by liquid chromatography coupled to electrospray ionization mass spectrometry and - for the main structures - by gas chromatography/mass spectrometry. Snail O-glycans are built from the four monosaccharide constituents: N-acetylgalactosamine, galactose, mannose and fucose. An additional modification is a methylation of the hexoses. The common trisaccharide core structure was determined in Arion lusitanicus to be N-acetylgalactosamine linked to the protein elongated by two 4-O-methylated galactose residues. Further elongations by methylated and unmethylated galactose and mannose residues and/or fucose are present. The typical snail O-glycan structures are different to those so far described. Similar to snail N-glycan structures they display methylated hexose residues.
Assuntos
Polissacarídeos/química , Caramujos/metabolismo , Animais , Cromatografia Gasosa-Espectrometria de Massas , Glicosilação , Dados de Sequência Molecular , Proteínas/química , Proteínas/metabolismo , Trissacarídeos/química , Trissacarídeos/metabolismoRESUMO
The carcinogenic compound N-nitrososarcosine (NSAR) is found in foods and tobacco products, and its quantification is of great interest. Although the presence of two stereoisomers, E- and Z-NSAR, is well-known, individual investigation of the isomers has not been reported so far. The present study by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) reveals that (i) the mass spectrometric responses of the isomers differ by a factor of approximately two and (ii) the isomer ratio is unstable in freshly prepared standard solutions. As a consequence, NSAR concentrations determined by LC-ESI-MS/MS are biased if those facts are not taken into account. The method described here overcomes the difficulty of stereospecific response by adjusting the isomer ratio and was applied to 100 tobacco products and fully validated for moist and dry snuff reference materials showing expanded measurement uncertainties of ~20% and limits of quantification of ~20 ng/g.
RESUMO
The N- and O-glycans of Arianta arbustorum, Achatina fulica, Arion lusitanicus and Planorbarius corneus were analysed for their monosaccharide pattern by reversed-phase HPLC after labelling with 2-aminobenzoic acid or 3-methyl-1-phenyl-2-pyrazolin-5-one and by gas chromatography-mass spectrometry. Glucosamine, galactosamine, mannose, galactose, glucose, fucose and xylose were identified. Furthermore, three different methylated sugars were detected: 3-O-methyl-mannose and 3-O-methyl-galactose were confirmed to be a common snail feature; 4-O-methyl-galactose was detected for the first time in snails.
Assuntos
Monossacarídeos/química , Oxigênio/química , Polissacarídeos/química , Caramujos/química , Animais , Galactose/química , Espectrometria de Massas , Metilação , Nitrogênio/químicaRESUMO
Glycosylation plays an important role in several types of recognition processes associated with fertilisation and development, allergies, pathological events and cell death. Whereas the amino acid sequence of a protein is fixed by the DNA, the glycosylation abilities depend on enzymes and substrates currently present in the cell.During the last decades our knowledge on glycosylation - the structure of glycans as well as the corresponding biochemical pathways including the responsible enzymes - especially on glycans of mammalian origin increased enormously. The glycosylation capabilities of other species were under investigation only if their glycans were for any reason connected to human life (e.g. some recognition processes of pathogens or allergy on food or plant glycans) or if they were potent candidates for cell culture systems for the expression of therapeutic agents (some insect, yeast and plant cells). However, in the meantime there is an increasing interest also in invertebrate glycosylation.Snails in particular show a broad spectrum of glycosylation abilities within their N-glycosylation pattern. In one case this has been shown to be involved in an intermediate host - parasite recognition process. For other snail species, it was found that they share many structural elements of N-glycans with mammals, plants, insects or nematodes. Sometimes several of these elements are present within one single structure.Here we present an overview of the current knowledge of N-glycosylation of snails, the glycan structures and the corresponding enzymes involved in the biosynthetic glycosylation pathway.