1. Identification of Glycosylated Sites in Rapana Hemocyanin by Mass Spectrometry and Gene Sequence, and Their Antiviral Effect
- Author
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Bernhard Lieb, Jozef Van Beeumen, Aleksander Dolashki, Bart Devreese, Angel S. Galabov, Lubomira Nikolaeva-Glomb, Pavlina Dolashka-Angelova, Nina Heilen, Koen Sandra, Stefan Stevanovic, Wolfgang Voelter, Ludmila Velkova, Institute of Organic Chemistry, Bulgarian Academy of Sciences (BAS), Institute of Zoology, Johannes Gutenberg - Universität Mainz (JGU), Department of Immunology, Institute for Cell Biology, Eberhard Karls Universität Tübingen = Eberhard Karls University of Tuebingen, Laboratory of Protein Biochemistry and Biomolecular Engineering, Universiteit Gent = Ghent University [Belgium] (UGENT), Institut Stephan Angeloff, Réseau International des Instituts Pasteur (RIIP), Institute of Biochemistry, and This work was supported by a research grant by the Ministry of Sciences and Education through the grant: DAAD-17/2007 (VU-L-310/07) and TK01/496, DFG (Deutsche Forschungsge- meinschaft) (Germany), Fund for Scientific Research-Flanders (FWO-Vlaanderen) through project VS.011.06N and CNR (Italy).
- Subjects
Models, Molecular ,medicine.medical_treatment ,Gastropoda ,Pharmaceutical Science ,Peptide ,MESH: Amino Acid Sequence ,Exon ,MESH: Animals ,Peptide sequence ,Chromatography, High Pressure Liquid ,chemistry.chemical_classification ,0303 health sciences ,biology ,Chemistry ,030302 biochemistry & molecular biology ,Glycopeptides ,Hemocyanin ,Glycopeptide ,3. Good health ,MESH: Hemocyanin ,[SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology ,Rapana ,Biochemistry ,Viruses ,MESH: Models, Molecular ,Biotechnology ,MESH: Antiviral Agents ,Spectrometry, Mass, Electrospray Ionization ,Molecular Sequence Data ,MESH: Sequence Alignment ,Biomedical Engineering ,Bioengineering ,Sequence alignment ,Antiviral Agents ,MESH: Spectrometry, Mass, Electrospray Ionization ,Virus ,MESH: Viruses ,03 medical and health sciences ,medicine ,Animals ,Amino Acid Sequence ,MESH: Chromatography, High Pressure Liquid ,MESH: Glycopeptides ,030304 developmental biology ,Pharmacology ,MESH: Gastropoda ,MESH: Molecular Sequence Data ,Organic Chemistry ,biology.organism_classification ,Hemocyanins ,Sequence Alignment - Abstract
International audience; Molluscan hemocyanins (Hcs) have recently received particular interest due to their significant immunostimulatory properties. This is mainly related to their high carbohydrate content and specific monosaccharide composition. We have now analyzed the oligosaccharides and the carbohydrate linkage sites of the Rapana venosa hemocyanin (RvH) using different approaches. We analyzed a number of glycopeptides by LC/ESI-MS/MS and identified the sugar chains and peptide sequences of 12 glycopeptides. Additionally, the potential carbohydrate linkage sites of 2 functional units, RvH-b and RvH-c, were determined by gene sequence analysis. Only RvH-c shows a potential N-glycosylation site. During this study, we discovered a highly conserved linker-intron, separating the coding exons of RVH-b and RvH-c. Following reports on antiviral properties from arthropod hemocyanin, we conducted a preliminary study of the antiviral activity of RvH and the functional units RvH-b and RvH-c. We show that the glycosylated FU RvH-c has antiviral properties against the respiratory syncytial virus (RSV), whereas native RvH and the nonglycosylated FU RvH-b have not. This is the first report of the fact that also molluscan hemocyanin functional units possess antiviral activity.
- Published
- 2009
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