1887

Abstract

Summary: Pseudomonas aeruginosa PML14e is a mutant strain, isolated from strain PML14 (Homma serotype I), that is resistant to all types of R-pyocins. PML14e completely lacked glucose and rhamnose as components of the lipopolysaccharide (LPS) outer core region. Whereas the O-polysaccharide attachment site on the LPS core was considered to be absent, PML14e was agglutinable with anti-serotype-I antibodies. The O-polysaccharide of PML14e was recovered in the supernatant after ultracentrifugation of the aqueous layer from a hot phenol/water extraction. Chromatographic behaviour and chemical analysis indicated that the PML14e O-polysaccharide was not linked to the lipid A. H-NMR spectroscopy indicated that the structure of the PML14e O-polysaccharide was the same as that of the O-polysaccharide from PML14. The above evidence indicated that the O-polysaccharide is expressed on the cell surface of the mutant strain PML14e as the lipid A-free form. To examine the nature of the cell surface, the accessibility of monoclonal antibodies (mAbs) against cell surface antigens was tested by enzyme-linked immunosorbent assay. An anti-lipid A mAb and an anti-outer-membrane protein mAb, the epitopes for which are considered to be exposed on rough strains, bound to a greater extent to the PML14e cells than to two other LPS-core-defective rough mutants, PML14b and PML14d. Whereas these mutants appeared to have lesser defects in the LPS core, they expressed less O-polysaccharide than PML14e. The results indicated that the epitopes exposed on rough strains, such as lipid A and outer-membrane proteins, were mainly hindered by covalently linked core oligosaccharide rather than by the O-polysaccharide chain.

Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-142-2-289
1996-02-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/142/2/mic-142-2-289.html?itemId=/content/journal/micro/10.1099/13500872-142-2-289&mimeType=html&fmt=ahah

References

  1. Beckmann I., Subbaiah T. V., Stocker B. A. D. 1964; Rough mutants of Salmonella typhimurium. 2. Serological and chemical investigations. Nature 201:1299–1301
    [Google Scholar]
  2. Berry D., Kropinski A. M. 1986; Effect of lipopolysaccharide mutations and temperature on plasmid transformation efficiency in Pseudomonas aeruginosa. Can J Microbiol 32:436–438
    [Google Scholar]
  3. Bhat R., Marx A., Galanos C., Conrad R. S. 1990; Structural studies of lipid A from Pseudomonas aeruginosa PAOl: Occurrence of 4-amino-4-deoxyarabinose. J Bacteriol 172:6631–6636
    [Google Scholar]
  4. Cloeckaert A., Zygmunt M. S., Nicolle J.-C., Dubray G., Limet J. N. 1992; O-chain expression in the rough Brucella melitensis strain B115: induction of O-polysaccharide-specific monoclonal antibodies and intracellular localization demonstrated by immuno-electron microscopy. J Gen Microbiol 138:1211–1219
    [Google Scholar]
  5. Coughlin R. T., Tonsager S., McGroarty E. J. 1983; Quantitation of met al cations bound to membranes and extracted lipopolysaccharide of Escherichia coli. Biochemistry 22:2002–2007
    [Google Scholar]
  6. Dasgupta T., deKievit T. R., Masoud H., Altman E., Richards J. C., Sadovskaya I., Speert D. P., Lam J. S. 1994; Characterization of lipopolysaccharide-deficient mutants of Pseudomonas aeruginosa derived from serotypes 03, 05, and 06. Infect Immun 62:809–817
    [Google Scholar]
  7. Dell A., Oates J., Lugowski C, Romanowska E., Kenne L., Lindberg B. 1984; The enterobacterial common-antigen, a cyclic polysaccharide. Carbohydr Res 133:95–104
    [Google Scholar]
  8. Dmitriev B. A., Kocharova N. A., Knirel Y. A., Shashkov A. S., Kochetkov N. K., Stanislavsky E. S., Mashilova G. M. 1982; Somatic antigens of Pseudomonas aeruginosa: the structure of the polysaccharide chain of Ps. aeruginosa 0:6 (Lanyi) lipopolysaccharide. Eur J Biochem 125:229–237
    [Google Scholar]
  9. Dubois M., Gilles K. A., Hamilton J. K., Rebers P. A., Smith F. 1956; Colorimetric method for determination of sugars and related substances. Anal Chem 28:350–356
    [Google Scholar]
  10. Galanos C., Luderitz O. 1975; Electrodialysis of lipopolysaccharide and their conversion to uniform salt forms. Eur J Biochem 54:603–610
    [Google Scholar]
  11. Goldman R. C., White D., Orskov F., Orskov I., Rick P. D., Lewis M. S., Bhattacharjee A. K., Leive L. 1982; A surface polysaccharide of Escherichia coli Olll contains O-antigen and inhibits agglutination of cells by O-antiserum. J Bacteriol 151:1210–1221
    [Google Scholar]
  12. Ito S., Kageyama M. 1970; Relationship between pyocins and a bacteriophage in Pseudomonas aeruginosa. J Gen Appl Microbiol 16:231–240
    [Google Scholar]
  13. Jann K., Jann B. 1983; The K antigens of Escherichia coli. Prog Allergy 33:53–79
    [Google Scholar]
  14. Jarrell K. F., Kropinski A. M. 1981a; Coliphage T7 receptors are present in Pseudomonas aeruginosa rough lipopolysaccharides. Biochem Biophys Res Commun 99:1185–1190
    [Google Scholar]
  15. Jarrell K. F., Kropinski A. M. 1981b; Pseudomonas aeruginosa bacteriophage ^PLS27-lipopolysaccharide interactions. J Virol 40:411–420
    [Google Scholar]
  16. Jarrell K. F., Kropinski A. M. 1981c; Isolation and characterization of a bacteriophage specific for the lipopolysaccharide of rough derivatives of Pseudomonas aeruginosa strain PAO. J Virol 38:529–538
    [Google Scholar]
  17. Karkhanis Y. D., Zeltner J. Y., Jackson J. J., Carlo D. J. 1978; A new and improved microassay to determine 2-keto-3-deoxy-octonate in lipopolysaccharide of gram-negative bacteria. Anal Biochem 85:595–601
    [Google Scholar]
  18. Knirel Y. A. 1990; Polysaccharide antigens of Pseudomonas aeruginosa. CRC Crit Rev Microbiol 17:273–304
    [Google Scholar]
  19. Koval S. F., Meadow P. M. 1977; The isolation and characterization of lipopolysaccharide-defective mutants of Pseudomonas aeruginosa PAC1. J Gen Microbiol 98:387–398
    [Google Scholar]
  20. Krallmann-Wenzel U., Schmidt G. 1994; A simple procedure to demonstrate the presence of the O-antigen capsule in entero-pathogenic Escherichia coli. Zentralbl Bakt 281:140–145
    [Google Scholar]
  21. Kuhn H.-M., Meier U., Mayer H. 1984; ECA, the enterobacterial common antigen - long neglected by microbiologists. Forum Mikrobiol 7:274–285 in German
    [Google Scholar]
  22. Liu P. V., Matsumoto H., Kusama H., Bergan T. 1983; Survey of heat-stable major somatic antigens of Pseudomonas aeruginosa. Int J Syst Bacteriol 33:256–264
    [Google Scholar]
  23. MacLachlan P. R., Keenleyside W. J., Dodgson C., Whitfield C. 1993; Formation of the K30 (group I) capsule in Escherichia coli 09: K30 does not require attachment to lipopolysaccharide lipid A-core. J Bacteriol 175:7515–7522
    [Google Scholar]
  24. Meadow P. M., Wells P. L. 1978; Receptor sites for R-type pyocins and bacteriophage E79 in the core part of the lipopolysaccharide of Pseudomonas aeruginosa PAC1. J Gen Microbiol 108:339–343
    [Google Scholar]
  25. Nikaido H. 1969; Biosynthesis of cell wall lipopolysaccharide in gram-negative enteric bacteria. Adv Ensymol 31:77–124
    [Google Scholar]
  26. Peterson A. A., McGroarty E. J. 1985; High-molecular-weight components in lipopolysaccharides of Salmonella typhimurium, Salmonella minnesota, and Escherichia coli. J Bacteriol 162:738–745
    [Google Scholar]
  27. Rowe P. S. N., Meadow P. M. 1983; Structure of the core oligosaccharide from the lipopolysaccharide of Pseudomonas aeruginosa PAC1R and its defective mutants. Eur J Biochem 132:329–337
    [Google Scholar]
  28. Seid R. C. Jr, Kopecko D. J., Sadoff J. C., Schneider H., Baron L. S., Formal S. B. 1984; Unusual lipopolysaccharide antigens of a Salmonella typhi oral vaccine strain expressing the Shigella sonnei form I antigen. J Biol Chem 259:9028–9034
    [Google Scholar]
  29. Shinomiya T., Shiga S. 1979; Bactericidal activity of the tail of Pseudomonas aeruginosa bacteriophage PS17. J Virol 32:958–967
    [Google Scholar]
  30. Suzuki N. 1988; Studies of endotoxic O-antigens from Pseudomonas aeruginosa. Seikagaku 60:1019–1046 in Japanese
    [Google Scholar]
  31. Temple G. S., Ayling P. D., Wilkinson S. G. 1986; Isolation and characterization of a lipopolysaccharide-specific bacteriophage of Pseudomonas aeruginosa. Microbios 45:81–91
    [Google Scholar]
  32. Tsuji A., Kinoshita T., Hoshino M. 1969; Microdetermination of hexosamines. Chem Pharm Bull 17:217–218
    [Google Scholar]
  33. Uchida K., Mizushima S. 1987; A simple method for isolation of lipopolysaccharides from Pseudomonas aeruginosa and some other bacterial strains. Agric Biol Chem 51:3107–3114
    [Google Scholar]
  34. Westphal O., Jann K. 1965; Bacterial lipopolysaccharides: extraction with phenol-water and further applications of the procedure. Methods Carbohydr Chem 5:83–91
    [Google Scholar]
  35. Wilkinson S. G. 1983; Composition and structure of lipopolysaccharides from Pseudomonas aeruginosa. Rev Infect Dis 5:S941–S949
    [Google Scholar]
  36. Yokota S. 1995; Identification of outer membrane proteins as target antigens of Pseudomonas aeruginosa Homma serotype M. Clin Diagn Lab Immunol 2:747–752
    [Google Scholar]
  37. Yokota S., Ochi H., Ohtsuka H., Kato M., Noguchi H. 1989; Heterogeneity of the l-rhamnose residue in the outer core of Pseudomonas aeruginosa lipopolysaccharide, characterized by using human monoclonal antibodies. Infect Immun 57:1691–1696
    [Google Scholar]
  38. Yokota S., Terashima M., Chiba J., Noguchi H. 1992; Variable cross-reactivity of Pseudomonas aeruginosa lipopolysaccharide-core-specific monoclonal antibodies and its possible relationship with serotype. J Gen Microbiol 138:289–296
    [Google Scholar]
  39. Yokota S., Hayashi T., Matsumoto H. 1994; Identification of the lipopolysaccharide core region as the receptor site for a cytotoxin-converting phage, φCTX, of Pseudomonas aeruginosa. J Bacteriol 176:5262–5269
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-142-2-289
Loading
/content/journal/micro/10.1099/13500872-142-2-289
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error