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Characterization of the cell surface glycolipid from Spirochaeta aurantia

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Abstract

Spirochaeta aurantia is a free-living saprophytic spirochete that grows easily in simple laboratory media, and thus can be used as a model for the investigation of surface carbohydrate structures in spirochetae, which are normally not available in sufficient amounts. Freeze-substitution electron microscopy indicated the presence of a capsule-like material projecting from the surface of S. aurantia. Extraction of cells gave two major glycolipids, the one with a higher molecular mass glycolipid was designated large glycolipid A (LGLA). LGLA contained small amount of branched and unsaturated O-linked fatty acids, l-rhamnose, l-fucose, d-xylose, d-mannose, d-glucosamine, d-glycero-d-gluco-heptose (DDglcHep), d-glycero-d-manno-heptose (DDHep), and a novel branched tetradeoxydecose monosaccharide, which we proposed to call aurantose (Aur). The carbohydrate structure of LGLA was extremely complex and consisted of the repeating units built of 11 monosaccharides, arrangement of nine of them was determined as:

$$\matrix {{\quad \quad \quad \quad \quad {\text{ - [ - 3 - }}\beta {\text{ - DDglcHep - }}3{\text{ - }}\beta {\text{ - D - GlcNAc - 2 - }}\beta {\text{ - D - Man - ] - }}}} \\ {{\quad \quad \quad \quad \quad \quad \quad \quad \quad \quad \quad \quad \quad \quad \quad \quad \quad \quad \quad \quad \quad |}} \\ {{\alpha {\text{ - Aur - 3 - }}\beta {\text{ - L - Rha - 4 - }}\beta {\text{ - D - Xyl - 4 - }}\alpha {\text{ - L - Fuc - 3 - }}\beta {\text{ - DDHep - 4}}}} \\ {{\quad \quad \quad \quad \quad \quad \quad \quad \quad |}} \\ {{\alpha {\text{ - L - Rha - 3}}}} \ $$

which wasdeduced from the NMR and chemical data on the LGLA and its fragments, obtained by various degradations. Tentative position of two remaining sugars is proposed. LGLA was negative for gelation of Limulus amebocyte lysate, did not contain lipid A, and was unable to activate any known Toll-like receptors.

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Acknowledgements

This work was supported by a Natural Sciences and Engineering Research Council of Canada Discovery grant to A.M.K. Authors are very indebted to Dr. Cristina De Castro, Università di Napoli “Federico II", for the identification of paratose and to Ms. Diane Moyles at the University of Guelph for freeze substitution electron microscopy.

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Authors and Affiliations

  1. Department of Biotechnology, Lund University, Lund, Sweden, 22100

    Catherine J. Paul

  2. Department of Microbiology, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA

    Elizabeth A. Lyle & Richard I. Tapping

  3. Department of Molecular & Cellular Biology, University of Guelph, Guelph, Ontario, Canada, N1G 2W1

    Terry J. Beveridge

  4. Department of Microbiology and Immunology, Queen’s University, Kingston, Ontario, Canada, K7L 3N6

    Andrew M. Kropinski

  5. Institute for Biological Sciences, National Research Council of Canada, 100 Sussex Dr, Ottawa, Ontario, Canada, K1A 0R6

    Evgeny Vinogradov

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  1. Catherine J. Paul
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Correspondence to Evgeny Vinogradov.

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Paul, C.J., Lyle, E.A., Beveridge, T.J. et al. Characterization of the cell surface glycolipid from Spirochaeta aurantia . Glycoconj J 26, 1097–1108 (2009). https://doi.org/10.1007/s10719-009-9230-4

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