1887

Abstract

The complete structures of LPS core types 1 and 2 from have been described by other authors. They are characterized by a lack of phosphoryl residues, but they contain galacturonic acid (GalA) residues, which contribute to the necessary negative charges. The presence of a capsule was determined in core-LPS non-polar mutants from strains 52145 (O1 : K2), DL1 (O1 : K1) and C3 (O8 : K66). O-antigen ligase () mutants produced a capsule. Core mutants containing the GalA residues were capsulated, while those lacking the residues were non capsulated. Since the proteins involved in the transfer of GalA (WabG) and glucosamine residues (WabH) are known, the chemical basis of the capsular-K2–cell-surface association was studied. Phenol/water extracts from 52145Δ and 52145Δ mutants, but not those from from 52145Δ mutant, contained both LPS and capsular polysaccharide, even after hydrophobic chromatography. The two polysaccharides were dissociated by gel-filtration chromatography, eluting with detergent and metal-ion chelators. From these results, it is concluded that the K2 capsular polysaccharide is associated by an ionic interaction to the LPS through the negative charge provided by the carboxyl groups of the GalA residues.

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2006-06-01
2019-10-14
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References

  1. Albertí, S., Rodríquez-Quiñones, F., Schirmer, T., Rummel, G., Tomás, J. M., Rosenbusch, J. P. & Benedí, V. J. ( 1995; ). A porin from Klebsiella pneumoniae: sequence homology, three-dimensional model, and complement binding. Infect Immun 63, 903–910.
    [Google Scholar]
  2. Arakawa, Y., Wacharotayankun, R., Nagatsuka, T., Ito, H., Kato, N. & Otha, M. ( 1995; ). Genomic organization of the Klebsiella pneumoniae cps region responsible for serotype K2 capsular polysaccharide synthesis in virulent strain Chedid. J Bacteriol 177, 1788–1796.
    [Google Scholar]
  3. Camprubí, S., Smith, M. A., Tomás, J. M. & Williams, P. ( 1992; ). Modulation of surface antigen expression by Klebsiella pneumoniae in response to growth environment. Microb Pathog 13, 145–155.[CrossRef]
    [Google Scholar]
  4. Climent, N., Ferrer, S., Rubires, X., Merino, S., Tomás, J. M. & Regué, M. ( 1997; ). Molecular characterization of a 17-kDa outer-membrane protein from Klebsiella pneumoniae. Res Microbiol 148, 133–143.[CrossRef]
    [Google Scholar]
  5. Corsaro, M. M., de Castro, C., Naldi, T., Parrilli, M., Tomás, J. M & Regué, M. ( 2005; ). 1H and 13C-NMR characterisation and secondary structure of the K2 polysaccharide of Klebsiella pneumoniae strain 52145. Carbohydr Res 340, 2212–2217.[CrossRef]
    [Google Scholar]
  6. Cortés, G., Borrell, N., Aztorza, B., Gómez, C., Sauleda, J. & Albertí, S. ( 2002; ). Molecular analysis of the contribution of the capsular polysaccharide and the lipopolysaccharide O side chain to the virulence of Klebsiella pneumoniae in a murine model of pneumonia. Infect Immun 70, 2583–2590.[CrossRef]
    [Google Scholar]
  7. Emori, T. G. & Gaynes, R. P. ( 1993; ). An overview of nosocomial infections, including the role of the microbiology laboratory. Clin Microbiol Rev 6, 428–442.
    [Google Scholar]
  8. Frirdich, E., Vinogradov, E. & Whitfield, C. ( 2004; ). Biosynthesis of a novel 3-deoxy-d-manno-oct-ulosonic acid-containing outer core oligosaccharide in the lipopolysaccharide of Klebsiella pneumoniae. J Biol Chem 279, 27928–27940.[CrossRef]
    [Google Scholar]
  9. Galanos, C., Lüderitz, O. & Westphal, O. ( 1969; ). A new method for the extraction of R lipopolysaccharides. Eur J Biochem 9, 245–249.[CrossRef]
    [Google Scholar]
  10. Gotschlich, E. C., Fraser, B. A., Nishimura, O., Robbins, J. B. & Liu, T.-Y. ( 1981; ). Lipid on capsular polysaccharides of Gram-negative bacteria. J Biol Chem 256, 8915–8921.
    [Google Scholar]
  11. Hansen, D. S., Mestre, F., Albertí, S. & 7 other authors ( 1999; ). Klebsiella pneumoniae lipopolysaccharide O typing: revision of prototype strains and O-group distribution among clinical isolates from different sources and countries. J Clin Microbiol 37, 56–62.
    [Google Scholar]
  12. Helander, I. M., Kato, Y., Kilpelainen, I., Kostiainen, R., Lindner, B., Nummila, K., Sugiyama, T. & Yokochi, T. ( 1996; ). Characterization of lipopolysaccharides of polymyxin-resistant and polymyxin-sensitive Klebsiella pneumoniae O3. Eur J Biochem 237, 272–278.[CrossRef]
    [Google Scholar]
  13. Hernández-Allés, S., Albertí, S., Alvarez, D., Domenech-Sánchez, A., Martinez-Martinez, L., Gil, J., Tomás, J. M. & Benedí, V. J. ( 1999; ). Porin expression in clinical isolates of Klebsiella pneumoniae. Microbiology 145, 673–679.[CrossRef]
    [Google Scholar]
  14. Hervás, J. A., Alomar, A., Salvá, F., Reina, J. & Benedí, V. J. ( 1993; ). Neonatal sepsis and meningitis in Mallorca (Spain), 1977–1991. Clin Infect Dis 16, 719–724.[CrossRef]
    [Google Scholar]
  15. Hitchcock, P. J. & Brown, T. M. ( 1983; ). Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels. J Bacteriol 154, 269–277.
    [Google Scholar]
  16. Holst, O., Brade, L., Kosma, P. & Brade, H. ( 1991; ). Structure, serological specificity, and synthesis of artificial glycoconjugates representing the genus-specific lipopolysaccharide epitope of Chlamydia spp. J Bacteriol 173, 1862–1866.
    [Google Scholar]
  17. Holst, O., Broer, W., Thomas-Oates, J. E., Mamat, U. & Brade, H. ( 1993; ). Structural analysis of two oligosaccharide bisphosphates isolated from the lipopolysaccharide of a recombinant strain of Escherichia coli F515 (Re chemotype) expressing the genus-specific epitope of Chlamydia lipopolysaccharide. Eur J Biochem 214, 703–710.[CrossRef]
    [Google Scholar]
  18. Izquierdo, L., Abitiu, N., Coderch, N., Hita, B., Merino, S., Gavín, R., Tomás, J. M. & Regué, M. ( 2002; ). The inner-core lipopolysaccharide biosynthetic waaE gene: function and genetic distribution among some Enterobacteriaceae. Microbiology 148, 3485–3496.
    [Google Scholar]
  19. Izquierdo, L., Coderch, N., Piqué, N., Bedini, E., Corsaro, M. M., Merino, S., Fresno, S., Tomas, J. M. & Regué, M. ( 2003; ). The Klebsiella pneumoniae wabG gene: its role in the biosynthesis of the core lipopolysaccharide and virulence. J Bacteriol 185, 7213–7221.[CrossRef]
    [Google Scholar]
  20. Jann, K., Dengler, T. & Jann, B. ( 1992; ). Core-lipid A on the K40 polysaccharide of Escherichia coli O8 : K40 : H9, a representative of group I capsular polysaccharides. Zentralbl Bakteriol 276, 196–204.[CrossRef]
    [Google Scholar]
  21. Kelly, R. F., Severn, W. B., Richards, J. C., Perry, M. B., MacLean, L. L., Tomás, J. M., Merino, S. & Whitfield, C. ( 1993; ). Structural variation in the O-specific polysaccharides of Klebsiella pneumoniae serotype O1 and O8 lipopolysaccharide: evidence for clonal diversity in rfb genes. Mol Microbiol 10, 615–625.[CrossRef]
    [Google Scholar]
  22. Knirel, Y. A. & Kochetkov, N. K. ( 1994; ). The structures of lipopolysaccharides of Gram-negative bacteria. III. The structure of O-antigens: a review. Biochemistry 59, 1325–1383.
    [Google Scholar]
  23. Kuo, S.-C., Doelling, V. W., Graveline, J. F. & McCoy, D. W. ( 1985; ). Evidence for covalent attachment of phospholipid to the capsular polysaccharide of Haemophilus influenzae type b. J Bacteriol 163, 769–773.
    [Google Scholar]
  24. Lai, Y.-C., Peng, H.-L. & Chang, H.-Y. ( 2003; ). RmpA2, an activator of capsule biosynthesis in Klebsiella pneumoniae CG43, regulates K2 cps gene expression at the transcriptional level. J Bacteriol 185, 788–800.[CrossRef]
    [Google Scholar]
  25. Link, A. J., Phillips, D. & Church, G. M. ( 1997; ). Methods for generating precise deletions and insertions in the genome of wild-type Escherichia coli: application to open reading frame characterization. J Bacteriol 179, 6228–6237.
    [Google Scholar]
  26. Martínez-Martínez, L., Hernández-Allés, S., Albertí, S., Tomás, J. M., Benedí, V. J. & Jacoby, G. A. ( 1996; ). In vivo selection of porin-deficient mutants of Klebsiella pneumoniae with increased resistance to cefoxitin and expanded-spectrum-cephalosporins. Antimicrob Agents Chemother 40, 342–348 (comment).
    [Google Scholar]
  27. Miller, J. H. ( 1972; ). Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  28. Muck, A., Ramm, M. & Hamburger, M. ( 1999; ). Efficient method of highly purified lipopolysaccharides by hydrophobic interaction chromatography. J Chromatogr B Biomed Sci Appl 732, 39–46.[CrossRef]
    [Google Scholar]
  29. Nassif, X., Fournier, J. M., Arondel, J. & Sansonetti, P. J. ( 1989; ). Mucoid phenotype of Klebsiella pneumoniae is a plasmid-encoded virulence factor. Infect Immun 57, 546–552.
    [Google Scholar]
  30. Nikaido, H. & Vaara, M. ( 1985; ). Molecular basis of bacterial outer membrane permeability. Microbiol Rev 49, 1–32.
    [Google Scholar]
  31. Ørskov, I. & Ørskov, F. ( 1984; ). Serotyping of Klebsiella. In Methods in Microbiology, pp. 143–164. Edited by T. Bergan. London: Academic Press.
  32. Pradel, E. & Schnaitman, C. A. ( 1991; ). Effect of rfaH (sfrB) and temperature on expression of rfa genes of Escherichia coli K-12. J Bacteriol 173, 6428–6431.
    [Google Scholar]
  33. Rahn, A., Beis, K., Naismith, J. H. & Whitfield, C. ( 2003; ). A novel outer membrane protein, Wzi, is involved in surface assembly of the Escherichia coli K30 group 1 capsule. J Bacteriol 185, 5882–5890.[CrossRef]
    [Google Scholar]
  34. Regué, M., Climent, N., Abitiu, N., Coderch, N., Merino, S., Izquierdo, L., Altarriba, M. & Tomás, J. M. ( 2001; ). Genetic characterization of the Klebsiella pneumoniae waa gene cluster, involved in core lipopolysaccharide biosynthesis. J Bacteriol 183, 3564–3573.[CrossRef]
    [Google Scholar]
  35. Regué, M., Hita, B., Piqué, N., Izquierdo, L., Merino, S., Fresno, S., Benedí, V. J. & Tomás, J. M. ( 2004; ). A gene (uridinediphosphate galacturonate 4-epimerase) is essential for Klebsiella pneumoniae virulence. Infect Immun 72, 54–61.[CrossRef]
    [Google Scholar]
  36. Regué, M., Izquierdo, L., Fresno, S. & 7 other authors ( 2005a; ). The incorporations of glucosamine to enterobacterial core LPS: two enzymatic steps are required. J Biol Chem 280, 36648–36656.[CrossRef]
    [Google Scholar]
  37. Regué, M., Izquierdo, L., Fresno, S. & 7 other authors ( 2005b; ). A second outer-core region in Klebsiella pneumoniae lipopolysaccharide. J Bacteriol 187, 4198–4206.[CrossRef]
    [Google Scholar]
  38. Sambrook, J., Fritsch, E. F. & Maniatis, T. ( 1989; ). Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  39. Straus, D. C., Atkisson, D. L. & Garner, C. W. ( 1985; ). Importance of a lipopolysaccharide-containing extracellular toxic complex in infections produced by Klebsiella pneumoniae. Infect Immun 50, 787–795.
    [Google Scholar]
  40. Süsskind, M., Muller-Loennies, S., Nimmich, W., Brade, H. & Holst, O. ( 1995; ). Structural investigation on the carbohydrate backbone of the lipopolysaccharide from Klebsiella pneumoniae rough mutant R20/O1. Carbohydr Res 269, 1–7.[CrossRef]
    [Google Scholar]
  41. Tomás, J. M., Camprubí, S. & Williams, P. ( 1988; ). Surface exposure of the O-antigen in Klebsiella pneumoniae O1 : K1 serotype strains. Microb Pathog 5, 141–147.[CrossRef]
    [Google Scholar]
  42. Tsai, C. M. & Frasch, C. E. ( 1982; ). A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem 119, 115–119.[CrossRef]
    [Google Scholar]
  43. Vinogradov, E. & Perry, M. B. ( 2001; ). Structural analysis of the core region of the lipopolysaccharides from eight serotypes of Klebsiella pneumoniae. Carbohydr Res 335, 291–296.[CrossRef]
    [Google Scholar]
  44. Vinogradov, E., Frirdich, E., MacLean, L. L., Perry, M. B., Petersen, B. O., Duus, J. O. & Whitfield, C. ( 2002; ). Structures of lipopolysaccharides from Klebsiella pneumoniae. Eluicidation of the structure of the linkage region between core and polysaccharide O chain and identification of the residues at the non-reducing termini of the O chains. J Biol Chem 277, 25070–25081.[CrossRef]
    [Google Scholar]
  45. Westphal, O. & Jann, K. ( 1965; ). Bacterial lipopolysaccharide extraction with phenol–water and further application of the procedure. Methods Carbohydr Chem 5, 83–91.
    [Google Scholar]
  46. Williams, P. & Tomás, J. ( 1990; ). The pathogenicity of Klebsiella pneumoniae. Rev Med Microbiol 1, 196–204.
    [Google Scholar]
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