Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1978 Dec;22(3):908–918. doi: 10.1128/iai.22.3.908-918.1978

Isolation and characterization of a high-molecular-weight polysaccharide from the slime of Pseudomonas aeruginosa.

G B Pier, H F Sidberry, S Zolyomi, J C Sadoff
PMCID: PMC422243  PMID: 103840

Abstract

A procedure is described for isolating a high-molecular-weight polysaccharide (PS) from the slime of Pseudomonas aeruginosa immunotype 1. The resultant material, obtained from the void volume of a Sephadex G-100 column, was composed of carbohydrate and water. No lipopolysaccharide (LPS), 2-keto-3-deoxyoctonoate, heptose, phosphate, or protein was detectable, and nucleic acid contamination was generally below 1%. The carbohydrate composition of the PS was glucose, rhamnose, galactose, arabinose, and mannose. PS had a molecular weight of between 100,000 and 350,000 and did not disaggregate when chromatographed in the presence of sodium deoxycholate. An antigen immunologically indistinguishable from PS could be obtained from LPS by either acetic acid hydrolysis and column chromatography or by allowing solutions of LPS to stand at room temperature for 3 days. Some of this LPS-associated polysaccharide eluted as the void volume of a G-100 column but differed from PS by its lack of galactose and arabinose. LPS also contained an immunodeterminant not shared with PS that was detected by its stability to dilute alkali treatment (0.1 N NaOH, 37 degrees C, 2 h). PS was destroyed by alkali treatment. PS appeared to represent a form of LPS polysaccharide side chain that contains galactose and arabinose and is of a high molecular weight.

Full text

PDF
908

Images in this article

915Image
on p.915

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alms T. H., Bass J. A. Immunization against Pseudomonas aeruginosa. I. Induction of protection by an alcohol-precipitated fraction from the slime layer. J Infect Dis. 1967 Jun;117(3):249–256. doi: 10.1093/infdis/117.3.249. [DOI] [PubMed] [Google Scholar]
  2. Austrian R. Prevention of pneumococcal infection by immunization with capsular polysaccharides of Streptococcus pneumoniae: current status of polyvalent vaccines. J Infect Dis. 1977 Aug;136 (Suppl):S38–S42. doi: 10.1093/infdis/136.supplement.s38. [DOI] [PubMed] [Google Scholar]
  3. Baltimore R. S., Dobek A. S., Stark F. R., Artenstein M. S. Clinical and epidemiological correlates of Pseudomonas typing. J Infect Dis. 1974 Nov;130 (Suppl)(0):S53–S59. doi: 10.1093/infdis/130.supplement.s53. [DOI] [PubMed] [Google Scholar]
  4. Bartell P. F., Orr T. E., Chudio B. Purification and Chemical Composition of the Protective Slime Antigen of Pseudomonas aeruginosa. Infect Immun. 1970 Nov;2(5):543–548. doi: 10.1128/iai.2.5.543-548.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bartell P. F., Orr T. E., Lam G. K. Polysaccharide depolymerase associated with bacteriophage infection. J Bacteriol. 1966 Jul;92(1):56–62. doi: 10.1128/jb.92.1.56-62.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chambers R. E., Clamp J. R. An assessment of methanolysis and other factors used in the analysis of carbohydrate-containing materials. Biochem J. 1971 Dec;125(4):1009–1018. doi: 10.1042/bj1251009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chester I. R., Meadow P. M. Heterogeneity of the lipopolysaccharide from Pseudomonas aeruginosa. Eur J Biochem. 1975 Oct 15;58(2):273–282. doi: 10.1111/j.1432-1033.1975.tb02373.x. [DOI] [PubMed] [Google Scholar]
  8. Drewry D. T., Lomax J. A., Gray G. W., Wilkinson S. G. Studies of lipid A fractions from the lipopolysaccharides of Pseudomonas aeruginosa and Pseudomonas alcaligenes. Biochem J. 1973 Jul;133(3):563–572. doi: 10.1042/bj1330563. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fensom A. H., Meadow P. M. Evidence for two regions in the polysaccharide moiety of the lipopolysaccharide of Pseudomonas aeruginosa 8602. FEBS Lett. 1970 Jul 29;9(2):81–84. doi: 10.1016/0014-5793(70)80318-0. [DOI] [PubMed] [Google Scholar]
  10. Fisher M. W., Devlin H. B., Gnabasik F. J. New immunotype schema for Pseudomonas aeruginosa based on protective antigens. J Bacteriol. 1969 May;98(2):835–836. doi: 10.1128/jb.98.2.835-836.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. GERLACH E., DEUTICKE B. EINE EINFACHE METHODE ZUR MIKROBESTIMMUNG VON PHOSPHAT IN DER PAPIERCHROMATOGRAPHIE. Biochem Z. 1963 Jul 26;337:477–479. [PubMed] [Google Scholar]
  12. Gotschlich E. C., Goldschneider I., Artenstein M. S. Human immunity to the meningococcus. V. The effect of immunization with meningococcal group C polysaccharide on the carrier state. J Exp Med. 1969 Jun 1;129(6):1385–1395. doi: 10.1084/jem.129.6.1385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. HAYNES W. C. Pseudomonas aeruginosa--its characterization and identification. J Gen Microbiol. 1951 Nov;5(5 Suppl):939–950. doi: 10.1099/00221287-5-5-939. [DOI] [PubMed] [Google Scholar]
  14. Hanessian S., Regan W., Watson D., Haskell T. H. Isolation and characterization of antigenic components of a new heptavalent Pseudomonas vaccine. Nat New Biol. 1971 Feb 17;229(7):209–210. doi: 10.1038/newbio229209a0. [DOI] [PubMed] [Google Scholar]
  15. Horton D., Rodemeyer G., Haskell T. H. Analytical characterization of lipopolysaccharide antigens from seven strains of Pseudomonas aeruginosa. Carbohydr Res. 1977 May;55:35–47. doi: 10.1016/s0008-6215(00)84441-9. [DOI] [PubMed] [Google Scholar]
  16. Hämmerling U., Westphal O. Synthesis and use of O-stearoyl polysaccharides in passive hemagglutination and hemolysis. Eur J Biochem. 1967 Mar;1(1):46–50. doi: 10.1007/978-3-662-25813-2_9. [DOI] [PubMed] [Google Scholar]
  17. Janda J., Work E. A colorimetric estimation of lipopolysaccharides. FEBS Lett. 1971 Sep 1;16(4):343–345. doi: 10.1016/0014-5793(71)80386-1. [DOI] [PubMed] [Google Scholar]
  18. KABAT E. A., BEZER A. E. The effect of variation in molecular weight on the antigenicity of dextran in man. Arch Biochem Biophys. 1958 Dec;78(2):306–318. doi: 10.1016/0003-9861(58)90354-0. [DOI] [PubMed] [Google Scholar]
  19. LIU P. V., ABE Y., BATES J. L. The roles of various fractions of Pseudomonas aeruginosa in its pathogenesis. J Infect Dis. 1961 Mar-Apr;108:218–228. doi: 10.1093/infdis/108.2.218. [DOI] [PubMed] [Google Scholar]
  20. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  21. Sadoff J. C. Cell-wall structures of Pseudomonas aeruginosa with immunologic significance: a brief review. J Infect Dis. 1974 Nov;130 (Suppl)(0):S61–S64. doi: 10.1093/infdis/130.supplement.s61. [DOI] [PubMed] [Google Scholar]
  22. Smith D. H., Peter G., Ingram D. L., Harding A. L., Anderson P. Responses of children immunized with the capsular polysaccharide of Hemophilus influenzae, type b. Pediatrics. 1973 Nov;52(5):637–644. [PubMed] [Google Scholar]
  23. Van der Wall C. A. The team approach to productivity improvement and cost containment. Purch Adm. 1978 Apr;2(3):8, 22-3. [PubMed] [Google Scholar]
  24. Weiss E., Long J. L. Simplified method for the production of carbohydrate antigen from Neisseria meningitidis. Appl Microbiol. 1969 Nov;18(5):843–847. doi: 10.1128/am.18.5.843-847.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Zollinger W. D., Dalrymple J. M., Artenstein M. S. Analysis of parameters affecting the solid phase radioimmunoassay quantitation of antibody to meningococcal antigens. J Immunol. 1976 Nov;117(5 PT2):1788–1798. [PubMed] [Google Scholar]
  26. Zollinger W. D., Mandrell R. E. Outer-membrane protein and lipopolysaccharide serotyping of Neisseria meningitidis by inhibition of a solid-phase radioimmunoassay. Infect Immun. 1977 Nov;18(2):424–433. doi: 10.1128/iai.18.2.424-433.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES