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. 1996 Mar;178(6):1731–1741. doi: 10.1128/jb.178.6.1731-1741.1996

Structure of the core oligosaccharide in the serotype O8 lipopolysaccharide from Klebsiella pneumoniae.

W B Severn 1, R F Kelly 1, J C Richards 1, C Whitfield 1
PMCID: PMC177860  PMID: 8626303

Abstract

Two classes of mutants with O-antigen-deficient lipopolysaccharides were isolated from the serotype O8 reference strain, belonging to Klebsiella pneumoniae subspecies ozaenae. These mutants were selected by resistance to bacteriophage KO1-2, which recognizes and lyses strains with lipopolysaccharide molecules containing the D-galactan II O antigen. Strain RFK-11 contains a defect in O-antigen synthesis and has a complete core, including the attachment site for O antigen. This mutation is complemented by a plasmid carrying the rfb (O-antigen biosynthesis) gene cluster from the related K. pneumoniae serotype O1. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the lipopolysaccharide from strain RFK-9 has a mobility typical of deep-rough lipopolysaccharide. RFK-9 lipopolysaccharide lacks the attachment site for O antigen. Lipopolysaccharides from strains RFK-9 and RFK-11 were isolated, and their structures were determined by methylation analyses, muclear magnetic resonance spectroscopy, and mass spectroscopy. The deduced O8 core oligosaccharide includes the partial core structure reported for the K. pneumoniae subspecies pneumoniae serotype O1 lipopolysaccharide (M. Süsskind, S. Müller-Leonnies, W. Nimmich, H. Brade, and O. Holst, Carbohydr. Res. 269:C1-7, 1995), consistent with the possibility of a conserved core structure within the species. The core oligosaccharide differs from those of the genera Salmonella and Escherichia by the absence of a hexose-containing outer core, the lack of phosphate residues in the inner core, and the presence of galacturonic acid residues.

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Selected References

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  1. Basu S., Radziejewska-Lebrecht J., Mayer H. Lipopolysaccharide of Providencia rettgeri. Chemical studies and taxonomical implications. Arch Microbiol. 1986 Apr;144(3):213–218. doi: 10.1007/BF00410949. [DOI] [PubMed] [Google Scholar]
  2. Binotto J., MacLachlan P. R., Sanderson K. E. Electrotransformation in Salmonella typhimurium LT2. Can J Microbiol. 1991 Jun;37(6):474–477. doi: 10.1139/m91-078. [DOI] [PubMed] [Google Scholar]
  3. Bock K., Vinogradov E. V., Holst O., Brade H. Isolation and structural analysis of oligosaccharide phosphates containing the complete carbohydrate chain of the lipopolysaccharide from Vibrio cholerae strain H11 (non-O1). Eur J Biochem. 1994 Nov 1;225(3):1029–1039. doi: 10.1111/j.1432-1033.1994.1029b.x. [DOI] [PubMed] [Google Scholar]
  4. Bronner D., Clarke B. R., Whitfield C. Identification of an ATP-binding cassette transport system required for translocation of lipopolysaccharide O-antigen side-chains across the cytoplasmic membrane of Klebsiella pneumoniae serotype O1. Mol Microbiol. 1994 Nov;14(3):505–519. doi: 10.1111/j.1365-2958.1994.tb02185.x. [DOI] [PubMed] [Google Scholar]
  5. Carlson R. W., Garci F., Noel D., Hollingsworth R. The structures of the lipopolysaccharide core components from Rhizobium leguminosarum biovar phaseoli CE3 and two of its symbiotic mutants, CE109 and CE309. Carbohydr Res. 1989 Dec 21;195(1):101–110. doi: 10.1016/0008-6215(89)85092-x. [DOI] [PubMed] [Google Scholar]
  6. Carlson R. W., Hollingsworth R. L., Dazzo F. B. A core oligosaccharide component from the lipopolysaccharide of Rhizobium trifolii ANU843. Carbohydr Res. 1988 May 1;176(1):127–135. doi: 10.1016/0008-6215(88)84064-3. [DOI] [PubMed] [Google Scholar]
  7. Clarke B. R., Bronner D., Keenleyside W. J., Severn W. B., Richards J. C., Whitfield C. Role of Rfe and RfbF in the initiation of biosynthesis of D-galactan I, the lipopolysaccharide O antigen from Klebsiella pneumoniae serotype O1. J Bacteriol. 1995 Oct;177(19):5411–5418. doi: 10.1128/jb.177.19.5411-5418.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Clarke B. R., Whitfield C. Molecular cloning of the rfb region of Klebsiella pneumoniae serotype O1:K20: the rfb gene cluster is responsible for synthesis of the D-galactan I O polysaccharide. J Bacteriol. 1992 Jul;174(14):4614–4621. doi: 10.1128/jb.174.14.4614-4621.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Coleman W. G., Jr The rfaD gene codes for ADP-L-glycero-D-mannoheptose-6-epimerase. An enzyme required for lipopolysaccharide core biosynthesis. J Biol Chem. 1983 Feb 10;258(3):1985–1990. [PubMed] [Google Scholar]
  10. Di Padova F. E., Brade H., Barclay G. R., Poxton I. R., Liehl E., Schuetze E., Kocher H. P., Ramsay G., Schreier M. H., McClelland D. B. A broadly cross-protective monoclonal antibody binding to Escherichia coli and Salmonella lipopolysaccharides. Infect Immun. 1993 Sep;61(9):3863–3872. doi: 10.1128/iai.61.9.3863-3872.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Domenico P., Diedrich D. L., Straus D. C. Extracellular polysaccharide production by Klebsiella pneumoniae and its relationship to virulence. Can J Microbiol. 1985 May;31(5):472–478. doi: 10.1139/m85-088. [DOI] [PubMed] [Google Scholar]
  12. Ferris F. G., Beveridge T. J. Site specificity of metallic ion binding in Escherichia coli K-12 lipopolysaccharide. Can J Microbiol. 1986 Jan;32(1):52–55. doi: 10.1139/m86-010. [DOI] [PubMed] [Google Scholar]
  13. HAKOMORI S. A RAPID PERMETHYLATION OF GLYCOLIPID, AND POLYSACCHARIDE CATALYZED BY METHYLSULFINYL CARBANION IN DIMETHYL SULFOXIDE. J Biochem. 1964 Feb;55:205–208. [PubMed] [Google Scholar]
  14. Hitchcock P. J., Brown T. M. Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels. J Bacteriol. 1983 Apr;154(1):269–277. doi: 10.1128/jb.154.1.269-277.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Holst O., Zähringer U., Brade H., Zamojski A. Structural analysis of the heptose/hexose region of the lipopolysaccharide from Escherichia coli K-12 strain W3100. Carbohydr Res. 1991 Aug 20;215(2):323–335. doi: 10.1016/0008-6215(91)84031-9. [DOI] [PubMed] [Google Scholar]
  16. Johnson K. G., Perry M. B. Improved techniques for the preparation of bacterial lipopolysaccharides. Can J Microbiol. 1976 Jan;22(1):29–34. doi: 10.1139/m76-004. [DOI] [PubMed] [Google Scholar]
  17. Kelly R. F., Severn W. B., Richards J. C., Perry M. B., MacLean L. L., Tomás J. M., Merino S., Whitfield C. Structural variation in the O-specific polysaccharides of Klebsiella pneumoniae serotype O1 and O8 lipopolysaccharide: evidence for clonal diversity in rfb genes. Mol Microbiol. 1993 Nov;10(3):615–625. doi: 10.1111/j.1365-2958.1993.tb00933.x. [DOI] [PubMed] [Google Scholar]
  18. Kol O., Wieruszeski J. M., Strecker G., Fournet B., Zalisz R., Smets P. Structure of the O-specific polysaccharide chain of Klebsiella pneumoniae O1K2 (NCTC 5055) lipopolysaccharide. A complementary elucidation. Carbohydr Res. 1992 Dec 15;236:339–344. doi: 10.1016/0008-6215(92)85028-x. [DOI] [PubMed] [Google Scholar]
  19. Kumar A., Ernst R. R., Wüthrich K. A two-dimensional nuclear Overhauser enhancement (2D NOE) experiment for the elucidation of complete proton-proton cross-relaxation networks in biological macromolecules. Biochem Biophys Res Commun. 1980 Jul 16;95(1):1–6. doi: 10.1016/0006-291x(80)90695-6. [DOI] [PubMed] [Google Scholar]
  20. Laakso D. H., Homonylo M. K., Wilmot S. J., Whitfield C. Transfer and expression of the genetic determinants for O and K antigen synthesis in Escherichia coli O9:K(A)30 and Klebsiella sp. O1:K20, in Escherichia coli K12. Can J Microbiol. 1988 Aug;34(8):987–992. doi: 10.1139/m88-173. [DOI] [PubMed] [Google Scholar]
  21. Montgomerie J. Z. Epidemiology of Klebsiella and hospital-associated infections. Rev Infect Dis. 1979 Sep-Oct;1(5):736–753. doi: 10.1093/clinids/1.5.736. [DOI] [PubMed] [Google Scholar]
  22. Nimmich W., Korten G. Die chemische Zusammensetzung der Klebsiella-Lipopolysaccharide (O-Antigene) Pathol Microbiol (Basel) 1970;36(3):179–190. [PubMed] [Google Scholar]
  23. Radziejewska-Lebrecht J., Mayer H. The core region of Proteus mirabilis R110/1959 lipopolysaccharide. Eur J Biochem. 1989 Aug 15;183(3):573–581. doi: 10.1111/j.1432-1033.1989.tb21086.x. [DOI] [PubMed] [Google Scholar]
  24. Rance M., Sørensen O. W., Bodenhausen G., Wagner G., Ernst R. R., Wüthrich K. Improved spectral resolution in cosy 1H NMR spectra of proteins via double quantum filtering. Biochem Biophys Res Commun. 1983 Dec 16;117(2):479–485. doi: 10.1016/0006-291x(83)91225-1. [DOI] [PubMed] [Google Scholar]
  25. Schnaitman C. A., Klena J. D. Genetics of lipopolysaccharide biosynthesis in enteric bacteria. Microbiol Rev. 1993 Sep;57(3):655–682. doi: 10.1128/mr.57.3.655-682.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Strain S. M., Fesik S. W., Armitage I. M. Structure and metal-binding properties of lipopolysaccharides from heptoseless mutants of Escherichia coli studied by 13C and 31P nuclear magnetic resonance. J Biol Chem. 1983 Nov 25;258(22):13466–13477. [PubMed] [Google Scholar]
  27. Straus D. C., Atkisson D. L., Garner C. W. Importance of a lipopolysaccharide-containing extracellular toxic complex in infections produced by Klebsiella pneumoniae. Infect Immun. 1985 Dec;50(3):787–795. doi: 10.1128/iai.50.3.787-795.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Straus D. C. Production of an extracellular toxic complex by various strains of Klebsiella pneumoniae. Infect Immun. 1987 Jan;55(1):44–48. doi: 10.1128/iai.55.1.44-48.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Süsskind M., Müller-Loennies S., Nimmich W., Brade H., Holst O. Structural investigation on the carbohydrate backbone of the lipopolysaccharide from Klebsiella pneumoniae rough mutant R20/O1-. Carbohydr Res. 1995 Apr 3;269(1):C1–C7. doi: 10.1016/0008-6215(95)00002-b. [DOI] [PubMed] [Google Scholar]
  30. Trautmann M., Vogt K., Hammack C., Cross A. S. A murine monoclonal antibody defines a unique epitope shared by Klebsiella lipopolysaccharides. Infect Immun. 1994 Apr;62(4):1282–1288. doi: 10.1128/iai.62.4.1282-1288.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tsai C. M., Frasch C. E. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem. 1982 Jan 1;119(1):115–119. doi: 10.1016/0003-2697(82)90673-x. [DOI] [PubMed] [Google Scholar]
  32. Vaara M. Agents that increase the permeability of the outer membrane. Microbiol Rev. 1992 Sep;56(3):395–411. doi: 10.1128/mr.56.3.395-411.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Whitfield C. Biosynthesis of lipopolysaccharide O antigens. Trends Microbiol. 1995 May;3(5):178–185. doi: 10.1016/s0966-842x(00)88917-9. [DOI] [PubMed] [Google Scholar]
  34. Whitfield C., Richards J. C., Perry M. B., Clarke B. R., MacLean L. L. Expression of two structurally distinct D-galactan O antigens in the lipopolysaccharide of Klebsiella pneumoniae serotype O1. J Bacteriol. 1991 Feb;173(4):1420–1431. doi: 10.1128/jb.173.4.1420-1431.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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