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Glycobiology Advance Access published online on June 2, 2005
Glycobiology, doi:10.1093/glycob/cwi084
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Published by Oxford University Press 2005.
Received November 8, 2004
Revised May 20, 2005
Accepted May 23, 2005

Article

Structures of the O-linked oligosaccharides of a complex glycoconjugate from Pseudallescheria boydii

Marcia R. Pinto 1, Philip A. J. Gorin 2, Robin Wait 3, Barbara Mulloy 4, and Eliana Barreto-Bergter 1

1 Instituto de Microbiologia, Universidade Federal do Rio de Janeiro, CCS, Bloco I, Ilha do Fundão, 21941-970, Rio de Janeiro, RJ, Brasil
2 Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Paraná (UFPR), 81531-990, Curitiba, PR, Brasil
3 Kennedy Institute of Rheumatology Division, Faculty of Medicine, Imperial College London, 1, Aspenlea Road, Hammersmith, London W6 8LH, UK
4 Laboratory for Molecular Structure, National Institute for Biological Standards and Control, Blanche Lane, South Mimms, Potters Bar, Herts. EN6 3QG UK


  Abstract

Non-reducing O-linked oligosaccharides were obtained from the peptidorhamnomannan of mycelia of Pseudallescheria boydii by alkaline {beta}- elimination under reducing conditions. They were separated by gel filtration chromatography to give three oligosaccharide fractions. The major oligosaccharide from fraction 1 was characterized by a combination of techniques including ESI MS/MS, MALDI MS, NMR, and methylation-GC-MS analysis. It was branched, with a principal chain of {alpha}-Rhap-(1->3)-{alpha}-Rhap-(1->3)-{alpha}-Manp-(1->2)-Man-ol substituted at O-6 of mannitol with an {alpha}-Glcp-(1->4)-{beta}-Galp group. Species containing one and two additional {alpha}-Glcp-(1->4) substituents in the rhamnose branch were also present. The major component of fraction 2 was a sub-structure of oligosaccharide-1, lacking a hexose from the Glc-Gal branch. Fraction 3 contained a mixture of smaller, unbranched, oligosaccharides. In hapten inhibition tests, fractions- 1 and -2 blocked the reaction between PRM and rabbit anti-P. boydii mycelium hyperimmune serum by about75%, while fraction-3 inhibited by about 55%.

Keywords: ELISA /ESI MS/MS/MALDI MS /methylation-GC-MS/Pseudallescheria boydii mycelia.
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Tracking the Emerging Human Pathogen Pseudallescheria boydii by Using Highly Specific Monoclonal Antibodies
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