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Purification and characterisation of lipoglycan macroamphiphiles from Propionibacterium acnes

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Abstract

Lipidated macroamphiphiles such as the lipoteichoic acids and mycobacterial lipoarabinomannans are cell envelope components of Gram-positive bacteria that have been extensively associated with the pathogenesis of disease. In order to study such associations, purification of these macroamphiphiles is essential for resolving their structures and diverse biological effects. We describe herein a method for purification of lipoglycan components from Propionibacterium acnes. This method uses the existing phenol-water extraction, followed by hydrophobic interaction chromatography and an additional purification step that utilises preparative electrophoresis for the separation of two lipoglycan components. Analysis of these lipoglycans revealed evidence for a lipid anchor based on fatty acids whilst the polysaccharide moiety contained significant amounts of mannose, glucose and galactose, together with an amino sugar suspected of being a diaminohexuronic acid. These latter components have been previously identified as components of the P. acnes cell wall polysaccharide. Consequently, it is proposed that there may be a relationship between the structures of these distinctive cell envelope polymers.

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References

  • Burton K. 1956. A study of the conditions and mechanism of the diphenylamine reaction for the colourimetric estimation of deoxyribonucleic acid. Biochem. J. 62: 315-323.

    Google Scholar 

  • Cantrell J.L. and Wheat R.W. 1979. Antitumor activity and lym-phoreticular stimulation properties of fractions isolated from Corynebacterium parvum. Cancer Res. 39: 3554-3564.

    Google Scholar 

  • Chatterjee D. and Khoo K.-H. 1998. Mycobacterial lipoarabi-nomannan: an extraordinary lipoheteroglycan with profound physiological effects. Glycobiology 8: 113-120.

    Google Scholar 

  • Chatterjee D., Roberts A.D., Lowell K., Brennan P.J. and Orme I.M. 1992. Structural basis of the capacity of lipoarabinomannan to induce secretion of tumour necrosis factor. Infect. Immun. 60: 1249-1253.

    Google Scholar 

  • Chen P.S., Toribara T.Y. and Warner H. 1956. Microdetermination of phosphorus. Anal. Chem. 28: 1756-1761.

    Google Scholar 

  • Cummins C.S. 1985. Distribution of 2,3-diamino-2,3-dideoxyglu-curonic acid in strains of Propionibacterium and other bacteria. Int. J. System. Bacteriol. 35: 411-416.

    Google Scholar 

  • Cummins C.S. and Johnson J.L. 1986. Genus I. Propionibacterium, Orla-Jensen 1909.. In: P.H.A. Sneath (ed.), Bergey's Manual of Systematic Bacteriology, pp. 1346-1353. Williams & Wilkins, London, UK.

    Google Scholar 

  • Cummins C.S. and White R.H. 1983. Isolation, identification, and synthesis of 2,3-diamino-2,3-dideoxyglucuronic acid: a component of Propionibacterium acnes cell wall polysaccharide. J. Bacteriol. 153: 1388-1393.

    Google Scholar 

  • Eady E.A. and Ingham E. 1999. Propionibacterium acnes-friend or foe. Rev. Med. Microbiol. 5: 163-173.

    Google Scholar 

  • Fischer W. 1991. One-step purification of bacterial lipid macroam-phiphiles by hydrophobic interaction chromatography. Anal. Biochem. 194: 353-358.

    Google Scholar 

  • Fischer W. 1993. Molecular analysis of lipid macroamphiphiles by hydrophobic interaction chromatography, exemplified with lipoteichoic acids. Anal. Biochem. 208: 49-56.

    Google Scholar 

  • Fischer W. 1994. Lipoteichoic acids and lipoglycans.. In: Ghuysen J.-M. and Hakenbeck R. (ed.), Bacterial Cell Wall, pp. 199-215. Elsevier Science, Amsterdam, The Netherlands.

    Google Scholar 

  • Fischer W. 1997. Pneumococcal lipoteichoic and teichoic acid. Microb. Drug Res. 3: 309-325.

    Google Scholar 

  • Fox J.D. and Robyt J.F. 1991. Miniturization of three carbohydrate analyses using a microsample plate reader. Anal. Biochem. 195: 93-96.

    Google Scholar 

  • Ginsburg I. 2002. Role of lipoteichoic acid in infection and inflammation. Lancet Infect. Dis. 2: 171-179.

    Google Scholar 

  • Hancock I. and Poxton I. 1988. Bacterial Cell Surface Techniques. John Wiley & Sons, Chichester, UK.

    Google Scholar 

  • Ingham E. 1999. The immunology of Propionibacterium acnes and acne. Curr. Op. Infect. Dis. 12: 191-197.

    Google Scholar 

  • Johnson J.L. and Cummins C.S. 1972. Cell wall composition and deoxyribonucleic acid similarities among the anaerobic coryne-forms, classical propionibacteria and strains of Arachnia propionica. J. Bacteriol. 109: 1047-1066.

    Google Scholar 

  • Kaur D, Lowary TL, Vissa VD, Crick DC and Brennan PJ. 2002. Characterization of the epitope of anti-lipoarabinomannan anti-bodies as the terminal hexaarabinofuranosyl motif of mycobacterial arabinans. Microbiology 148: 3049-3057.

    Google Scholar 

  • Kokeguchi S., Ohto H., Fukui K., Kato K., Shimizu H., Kurihara H., Kinoshita M., Nomura Y. and Murayama Y. 1985. Amphiphile antigen from Propionibacterium acnes: chemical composition, immunochemical properties and cytotoxicity on human gingival fibroblasts. J. Okayama Dent. Soc. 4: 89-102.

    Google Scholar 

  • Laemmli U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685.

    Google Scholar 

  • Mamat U., Seydel U., Grimmecke D., Holst O. and Rietschel E.T. 1999. Lipopolysaccharides. In: Comprehensive Natural Products Chemistry, pp. 179-239. Elservier Science, Amsterdam, The Netherlands.

    Google Scholar 

  • Mára M., Julák J., Bednár M., Ocenásková J., Miková Z. and Dohnalová A. 1994. The influence of Propionibacterium acnes (Corynebacterium parvum) fractions on immune response in vivo. Zentralbl. Bakteriol. 281: 549-555.

    Google Scholar 

  • Mordarska H. and Pas´ciak M. 1994. A simple method for the differentiation of Propionibacterium acnes and Propionibacterium propionicum. FEMS Microbiol. Letts. 123: 325-330.

    Google Scholar 

  • Moss C., Dowell V.R., Farshtchi D., Raines LJ. and Cherry W.B. 1969. Cultural characteristics and fatty acid composition of pro-pionibacteria. J. Bacteriol. 97: 561-57.

    Google Scholar 

  • Nagaoka M., Kamisango K., Fujii H., Uchikawa K., Sekikawa I. and Azuma I. 1985. Structure of acidic polysaccharide from cell wall of Propionibacterium acnes strain C7. J. Biochem. 97: 1669-1678.

    Google Scholar 

  • Nigou J., Gilleron M. and Puzo G. 2003. Lipoarabinomannans: from structure to biosynthesis. Biochimie 85: 153-166.

    Google Scholar 

  • Smith P.K., Krohn R.I., Hermanson G.T., Mallia A.K., Gartner F.H., Provenzano M.D., Fujimoto E.K., Goeke N.M., Olson B.J. and Klenk D.C. 1985. Measurement of protein using bicincho-ninic acid. Anal. Biochem. 150: 76-85.

    Google Scholar 

  • Stackebrandt E., Rainey F.A. and Ward-Rainey N.L. 1997. Proposal for a new hierarchic classification system, Actinobac-teria classis nov. Int. J. Syst. Bacteriol. 47: 479-491.

    Google Scholar 

  • Strohmeier G.R. and Fenton M.J. 1999. Roles of lipoarabinoman-nan in the pathogenesis of tuberculosis. Microb. Infect. 1: 709-717.

    Google Scholar 

  • Sutcliffe I.C. 2000. Characterisation of a lipomannan lipoglycan from the mycolic acid containing actinomycete Dietzia maris. Antonie van Leeuwenhoek 78: 195-201.

    Google Scholar 

  • Sutcliffe I.C. and Shaw N. 1989. An inositol-containing lipoman-nan from Propionibacterium freudenreichii. FEMS Microbiol. Letts. 59: 249-252.

    Google Scholar 

  • Sutcliffe I.C. and Shaw N. 1993. The phospholipids of Propioni-bacterium freudenreichii: absence of phosphatidylinositol man-nosides. System. Appl. Microbiol. 16: 9-12.

    Google Scholar 

  • Sutcliffe I.C. 1994. The lipoteichoic acids and lipoglycans of Gram-positive bacteria: a chemotaxanomic Perspective. System. Appl. Microbiol. 17: 467-480.

    Google Scholar 

  • Towbin H., Staehelin T. and Gordon J. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Nat. Acad. Sci. USA 76: 4350-4354.

    Google Scholar 

  • Tsai C.-M. and Frasch C.E. 1982. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal. Bio-chem. 119: 115-119.

    Google Scholar 

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Whale, G.A., Sutcliffe, I.C., Morrisson, A.R. et al. Purification and characterisation of lipoglycan macroamphiphiles from Propionibacterium acnes . Antonie Van Leeuwenhoek 86, 77–85 (2004). https://doi.org/10.1023/B:ANTO.0000024911.67625.27

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  • DOI: https://doi.org/10.1023/B:ANTO.0000024911.67625.27

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