1887

Abstract

Enterobacterial common antigen (ECA) is a polysaccharide present in all members of Enterobacteriaceae anchored either via phosphatidylglycerol (PG) or LPS to the outer leaflet of the outer membrane (ECA and ECA, respectively). Only the latter form is ECA-immunogenic. We previously demonstrated that O : 3 and its rough (O-specific polysaccharide-negative) mutants were ECA-immunogenic, suggesting that they contained ECA; however, it was not known which part of the LPS core region was involved in ECA binding. To address this, we used a set of three deep-rough LPS mutants for rabbit immunization. The polyvalent antisera obtained were: (i) analysed for the presence of anti-LPS and anti-ECA antibodies; (ii) treated with caprylic acid (CA) to precipitate IgM antibodies and protein aggregates; and (iii) adsorbed with live ECA-negative bacteria to obtain specific anti-ECA antisera. We demonstrated the presence of antibodies specific for both ECA and ECA in all antisera obtained. Both CA treatment and adsorption with ECA-negative bacteria efficiently removed anti-LPS antibodies, resulting in specific anti-ECA sera. The LPS of the ECA-positive deepest-rough mutant contained only lipid A and 3-deoxy---oct-2-ulosonic acid (Kdo) residues of the inner core, suggesting that ECA was linked to the Kdo region of LPS in O : 3.

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2015-01-01
2021-08-02
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References

  1. Alexander C., Rietschel E. T. 2001; Bacterial lipopolysaccharides and innate immunity. J Endotoxin Res 7:167–202[PubMed]
    [Google Scholar]
  2. al-Hendy A., Toivanen P., Skurnik M. 1992; Lipopolysaccharide O side chain of Yersinia enterocolitica O : 3 is an essential virulence factor in an orally infected murine model. Infect Immun 60:870–875[PubMed]
    [Google Scholar]
  3. Bottone E. J. 1999; Yersinia enterocolitica: overview and epidemiologic correlates. Microbes Infect 1:323–333 [View Article][PubMed]
    [Google Scholar]
  4. Brade H., Galanos C., Lüderitz O. 1983; Differential determination of the 3-deoxy-d-mannooctulosonic acid residues in lipopolysaccharides of Salmonella minnesota rough mutants. Eur J Biochem 131:195–200 [View Article][PubMed]
    [Google Scholar]
  5. Castelli M. E., Véscovi E. G. 2011; The Rcs signal transduction pathway is triggered by enterobacterial common antigen structure alterations in Serratia marcescens. J Bacteriol 193:63–74 [View Article][PubMed]
    [Google Scholar]
  6. Chung H. S., Raetz C. R. H. 2010; Dioxygenases in Burkholderia ambifaria and Yersinia pestis that hydroxylate the outer Kdo unit of lipopolysaccharide. Proc Natl Acad Sci U S A 108:510–215 [View Article][PubMed]
    [Google Scholar]
  7. Duda K. A. 2007a; Immunochemical studies on lipopolysaccharides from R mutants Yersinia enterocolitica O : 3. PhD thesis, University of Silesia.
    [Google Scholar]
  8. Duda K. T. 2007b; Reactivity of polyvalent antisera against R mutants of Yersinia enterocolitica O : 3. PhD thesis, University of Silesia.
    [Google Scholar]
  9. Duda K. A., Lindner B., Brade H., Leimbach A., Brzuszkiewicz E., Dobrindt U., Holst O. 2011; The lipopolysaccharide of the mastitis isolate Escherichia coli strain 1303 comprises a novel O-antigen and the rare K-12 core type. Microbiology 157:1750–1760 [View Article][PubMed]
    [Google Scholar]
  10. Focà A., Liberto M. C., Quirino A., Matera G. 2012; Lipopolysaccharides: from Erinyes to Charites. Mediators Inflamm 2012:684274 [View Article][PubMed]
    [Google Scholar]
  11. Fredriksson-Ahomaa M., Korkeala H. 2003; Low occurrence of pathogenic Yersinia enterocolitica in clinical, food, and environmental samples: a methodological problem. Clin Microbiol Rev 16:220–229 [View Article][PubMed]
    [Google Scholar]
  12. Galanos C., Lüderitz O., Westphal O. 1969; A new method for the extraction of R lipopolysaccharides. Eur J Biochem 9:245–249 [View Article][PubMed]
    [Google Scholar]
  13. Gilbreath J. J., Colvocoresses Dodds J., Rick P. D., Soloski M. J., Merrell D. S., Metcalf E. S. 2012; Enterobacterial common antigen mutants of Salmonella enterica serovar Typhimurium establish a persistent infection and provide protection against subsequent lethal challenge. Infect Immun 80:441–450 [View Article][PubMed]
    [Google Scholar]
  14. Goverde R. L. J., Huis in’t Veld J. H., Kusters J. G., Mooi F. R. 1998; The psychrotrophic bacterium Yersinia enterocolitica requires expression of pnp, the gene for polynucleotide phosphorylase, for growth at low temperature (5°C). Mol Microbiol 28:555–569 [View Article][PubMed]
    [Google Scholar]
  15. Gozdziewicz T. K., Lugowski C., Lukasiewicz J. 2014; First evidence for a covalent linkage between enterobacterial common antigen and lipopolysaccharide in Shigella sonnei phase II ECALPS. J Biol Chem 289:2745–2754 [View Article][PubMed]
    [Google Scholar]
  16. Gronow S., Brabetz W., Brade H. 2000; Comparative functional characterization in vitro of heptosyltransferase I (WaaC) and II (WaaF) from Escherichia coli. Eur J Biochem 267:6602–6611 [View Article][PubMed]
    [Google Scholar]
  17. Hoffman J., Lindberg B., Brubaker R. R. 1980; Structural studies of the O-specific side-chains of the lipopolysaccharide from Yersinia enterocolitica Ye 128. Carbohydr Res 78:212–214 [View Article][PubMed]
    [Google Scholar]
  18. Kaca W., de Jongh-Leuvenink J., Zähringer U., Rietschel E. T., Brade H., Verhoef J., Sinnwell V. 1988; Isolation and chemical analysis of 7-O-(2-amino-2-deoxy-α-d-glucopyranosyl)-l-glycero-d-manno-heptose as a constituent of the lipopolysaccharides of the UDP-galactose epimerase-less mutant J-5 of Escherichia coli and Vibrio cholerae. Carbohydr Res 179:289–299 [View Article][PubMed]
    [Google Scholar]
  19. Kasperkiewicz K. 2002; ECA-immunogenicity of Yersinia enterocolitica rough mutants. PhD thesis, University of Silesia.
    [Google Scholar]
  20. Kuhn H.-M., Meier-Dieter U., Mayer H. 1988; ECA, the enterobacterial common antigen. FEMS Microbiol Rev 54:195–222 [View Article][PubMed]
    [Google Scholar]
  21. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685 [View Article][PubMed]
    [Google Scholar]
  22. Männel D., Mayer H. 1978; Isolation and chemical characterization of the enterobacterial common antigen. Eur J Biochem 86:361–370 [View Article][PubMed]
    [Google Scholar]
  23. McArthur F., Andersson C. E., Loutet S., Mowbray S. L., Valvano M. A. 2005; Functional analysis of the glycero-manno-heptose 7-phosphate kinase domain from the bifunctional HldE protein, which is involved in ADP-l-glycero-d-manno-heptose biosynthesis. J Bacteriol 187:5292–5300 [View Article][PubMed]
    [Google Scholar]
  24. Meredith T. C., Aggarwal P., Mamat U., Lindner B., Woodard R. W. 2006; Redefining the requisite lipopolysaccharide structure in Escherichia coli. ACS Chem Biol 1:33–42 [View Article][PubMed]
    [Google Scholar]
  25. Michiels T., Cornelis G. R. 1991; Secretion of hybrid proteins by the Yersinia Yop export system. J Bacteriol 173:1677–1685[PubMed]
    [Google Scholar]
  26. Muszyński A., Rabsztyn K., Knapska K., Duda K. A., Duda-Grychtoł K. T., Kasperkiewicz K., Radziejewska-Lebrecht J., Holst O., Skurnik M. 2013; Enterobacterial common antigen and O-specific polysaccharide coexist in the lipopolysaccharide of Yersinia enterocolitica serotype O : 3. Microbiology 159:1782–1793 [View Article][PubMed]
    [Google Scholar]
  27. Pinta E., Duda K. A., Hanuszkiewicz A., Kaczyński Z., Lindner B., Miller W. L., Hyytiäinen H., Vogel Ch., Borowski S.& other authors ( 2009; Identification and role of a 6-deoxy-4-keto-hexosamine in the lipopolysaccharide outer core of Yersinia enterocolitica serotype O : 3. Chemistry 15:9747–9754 [View Article][PubMed]
    [Google Scholar]
  28. Pinta E., Li Z., Batzilla J., Pajunen M., Kasanen T., Rabsztyn K., Rakin A., Skurnik M. 2012; Identification of three oligo-/polysaccharide-specific ligases in Yersinia enterocolitica. Mol Microbiol 83:125–136 [View Article][PubMed]
    [Google Scholar]
  29. Pupek M. 2009; The antibodies as reagents in immunochemistry. In Immunochemistry in medical biology pp. 18–54 Edited by Ka˛tnik-Prastowska I. Warsaw: Polish Scientific Publishers [in Polish];
    [Google Scholar]
  30. Rabsztyn K., Kasperkiewicz K., Duda K. A., Li C.-M., Łukasik M., Radziejewska-Lebrecht J., Skurnik M. 2011; Characterization of anti-ECA antibodies in rabbit antiserum against rough Yersinia enterocolitica O : 3. Biochemistry (Mosc) 76:832–839 [View Article][PubMed]
    [Google Scholar]
  31. Radziejewska-Lebrecht J., Shashkov A. S., Stroobant V., Wartenberg K., Warth Ch., Mayer H. 1994; The inner core region of Yersinia enterocolitica Ye75R (O : 3) lipopolysaccharide. Eur J Biochem 221:343–351 [View Article][PubMed]
    [Google Scholar]
  32. Radziejewska-Lebrecht J., Skurnik M., Shashkov A. S., Brade L., Różalski A., Bartodziejska B., Mayer H. 1998; Immunochemical studies on R mutants of Yersinia enterocolitica O : 3. Acta Biochim Pol 45:1011–1019[PubMed]
    [Google Scholar]
  33. Radziejewska-Lebrecht J., Kasperkiewicz K., Skurnik M., Brade L., Steinmetz I., Świerzko A. S., Muszyński A. 2003; ECA-antibodies in antisera against R mutants of Yersinia enterocolitica O : 3. In The Genus Yersinia: Entering the Functional Genomic Era pp. 215–218 Edited by Skurnik M., Granfors K., Bengoechea J. A. New York: Kluwer Academic/Plenum Publishers;
    [Google Scholar]
  34. Seltmann G., Holst O. 2001; The outer membrane of the Gram-negative bacteria and their components. In The Bacterial Cell Wall pp. 31–66 Edited by Seltman G., Holst O. Berlin: Springer;
    [Google Scholar]
  35. Shashkov A. S., Radziejewska-Lebrecht J., Kochanowski H., Mayer H. 1995; The chemical structure of the outer core region of the Yersinia enterocolitica O : 3 lipopolysaccharide. In Abstract BO17, 8th European Carbohydrate Symposium Sevilla, Spain:
    [Google Scholar]
  36. Skurnik M. 1984; Lack of correlation between the presence of plasmids and fimbriae in Yersinia enterocolitica and Yersinia pseudotuberculosis. J Appl Bacteriol 56:355–363 [View Article][PubMed]
    [Google Scholar]
  37. Skurnik M., Venho R., Toivanen P., Al-Hendy A. 1995; A novel locus of Yersinia enterocolitica serotype O : 3 involved in lipopolysaccharide outer core biosynthesis. Mol Microbiol 17:575–594 [View Article][PubMed]
    [Google Scholar]
  38. Skurnik M., Venho R., Bengoechea J.-A., Moriyón I. 1999; The lipopolysaccharide outer core of Yersinia enterocolitica serotype O : 3 is required for virulence and plays a role in outer membrane integrity. Mol Microbiol 31:1443–1462 [View Article][PubMed]
    [Google Scholar]
  39. Swierzko A., Brade L., Paulsen H., Brade H. 1993; Specificity of rabbit antisera against the rough lipopolysaccharide of Salmonella minnesota R4 (chemotype Rd2P-). Infect Immun 61:3216–3221[PubMed]
    [Google Scholar]
  40. Swierzko A., Brade L., Brabetz W., Zych K., Paulsen H., Brade H. 1994; Monoclonal antibodies against the heptose region of enterobacterial lipopolysaccharides. Innate Immun 1:38–44 [View Article]
    [Google Scholar]
  41. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A 76:4350–4354 [View Article][PubMed]
    [Google Scholar]
  42. Wartenberg K., Knapp W., Ahamed N. M., Widemann C., Mayer H. 1983; Temperature-dependent changes in the sugar and fatty acid composition of lipopolysaccharides from Yersinia enterocolitica strains. Zentralbl Bakteriol Mikrobiol Hyg [A] 253:523–530[PubMed]
    [Google Scholar]
  43. Wollenweber H. W., Rietschel E. T. 1990; Analysis of lipopolysaccharide (lipid A) fatty acids. J Microbiol Methods 11:195–211 [View Article]
    [Google Scholar]
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