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Journal of Botany
Volume 2009 (2009), Article ID 560394, 12 pages
Research Article

Partial Characterization of a Vicilin-Like Glycoprotein from Seeds of Flowering Tobacco (Nicotiana sylvestris)

1Center for Glycosciences and Technology, Biodesign Institute at Arizona State University, 1001 S McAllister Avenue, Tempe, AZ 85287, USA
2National Centre for Biomedical Engineering Science, National University of Ireland, Galway, Ireland
3Bio5 Institute for Collaborative Bioresearch, The University of Arizona, Tucson, AZ 85721, USA
4Department of Chemistry and Biomolecular Sciences, Macquarie University, North Ryde NSW 2109, Australia
5Australian Proteome Analysis Facility, Macquarie University, North Ryde NSW 2109, Australia

Received 5 February 2009; Accepted 31 March 2009

Academic Editor: Andrew Wood

Copyright © 2009 Jared Q. Gerlach et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


A vicilin-like glycoprotein from the seeds of Nicotiana sylvestris, flowering tobacco, has been identified using nanoLC/ESI-MS/MS. Sequences from a fragment of protein demonstrated homology with vicilins from other members of the Solanaceae family, notably potato (Solanum demissum). Reducing and nonreducing SDS-PAGE analyses of the identified protein indicated that fragments resulting from in situ proteolytic processing are joined by intrachain disulphide bonds. Staining with Con A lectin was specifically inhibited by mannose suggested the presence of -linked glycosylation which was confirmed by carbohydrate compositional analysis of PVDF-bound protein subunits. HPAEC-PAD analysis of the monosaccharides released from the glycoprotein by acid hydrolysis revealed glucosamine and mannose. -acetylglucosamine termination of attached oligosaccharides was further verified by inhibitable WGA lectin staining. Immunostaining of PVDF-bound N. sylvestris proteins with antibodies against G. max total protein demonstrated cross-staining at masses corresponding to fragments from the proteolytically processed protein subunits.