Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1979 Mar;23(3):845–857. doi: 10.1128/iai.23.3.845-857.1979

Isolation, characterization, and biological properties of an endotoxin-like material from the gram-positive organism Listeria monocytogenes.

H Wexler, J D Oppenheim
PMCID: PMC414241  PMID: 110684

Abstract

The bacterial component responsible for the induction of transient cold agglutinin syndrome in rabbits after intravenous injection of heat-killed Listeria monocytogenes type 4B has been purified and biologically and chemically characterized. A purified immunoglobulin M cold agglutinin was prepared from high-titer sera resulting from the immunization of rabbits with heat-killed L. monocytogenes type 4B and was subsequently used to monitor the purification of the bacterial component responsible for its induction. The bacterial component was isolated from a hot phenol-water extract of lyophilized L. monocytogenes type 4B by multiple molecular sieve chromatography. Upon chemical analysis the purified material was found to be strikingly similar in chemical composition to gram-negative lipopolysaccharide endotoxins. The material contained 15% total fatty acid (of which 50% was beta-hydroxymyristic acid), 40 to 45% neutral sugar (glucose, galactose, and rhamnose), 11.5% amino sugar, 12% uronic acid, 2.5% 2-keto-3-deoxyoctonic acid, 2% heptose, 0.87% phosphorus, and 1.6% amino acid, thereby accounting for 85 to 90% of the weight of the component. Electron micrographs of the purified material were similar to those of lipopolysaccharide preparations from gram-negative organisms. The purified material exist in aqueous solutions as large aggregates, but can be dissociated into a single smaller subunit (3.1S) by dialysis against sodium dodecyl sulfate buffer. The listerial component was toxic and pyrogenic to rabbits, producing symptoms typical of gram-negative endotoxins. Activity in the limulus lysate gelation assay and in the carbocyanine dye assay provides a further link of this material with classical gram-negative endotoxins.

Full text

PDF
845

Images in this article

850Image
on p.850
851Image
on p.851
851Image
on p.851

Selected References

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

  1. Anacker R. L., Bickel W. D., Haskins W. T., Milner K. C., Ribi E., Rudbach J. A. Frequency of occurrence of native hapten among enterobacterial species. J Bacteriol. 1966 Apr;91(4):1427–1433. doi: 10.1128/jb.91.4.1427-1433.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  3. Blumenkrantz N., Asboe-Hansen G. An assay for total hexosamine and a differential assay for glucosamine and galactosamine. Clin Biochem. 1976 Dec;9(6):269–274. doi: 10.1016/s0009-9120(76)80075-6. [DOI] [PubMed] [Google Scholar]
  4. COSTEA N., YAKULIS V., HELLER P. EXPERIMENTAL PRODUCTION OF COLD AGGLUTININS IN RABBITS. Blood. 1965 Sep;26:323–340. [PubMed] [Google Scholar]
  5. Conklin A. R., Jr, Siddique I. H. Certain chemical and biological properties of phenol extracts from Listeria monocytogenes. Am J Vet Res. 1976 Nov;37(11):1331–1335. [PubMed] [Google Scholar]
  6. Costea N., Yakulis V. J., Heller P. Inhibition of cold agglutinins (anti-I) by M. pneumoniae antigens. Proc Soc Exp Biol Med. 1972 Feb;139(2):476–479. doi: 10.3181/00379727-139-36167. [DOI] [PubMed] [Google Scholar]
  7. Costea N., Yakulis V. J., Heller P. The mechanism of indction of cold agglutinins by mycoplasma pneumoniae. J Immunol. 1971 Mar;106(3):598–604. [PubMed] [Google Scholar]
  8. DISCHE Z. New color reactions for determination of sugars in polysaccharides. Methods Biochem Anal. 1955;2:313–358. doi: 10.1002/9780470110188.ch11. [DOI] [PubMed] [Google Scholar]
  9. Duncombe W. G. The colorimetric micro-determination of long-chain fatty acids. Biochem J. 1963 Jul;88(1):7–10. doi: 10.1042/bj0880007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  11. Gray M. L., Killinger A. H. Listeria monocytogenes and listeric infections. Bacteriol Rev. 1966 Jun;30(2):309–382. doi: 10.1128/br.30.2.309-382.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Jackson S. W., Zey P. N. Ultrastructure of lipopolysaccharide isolated from Treponema pallidum. J Bacteriol. 1973 May;114(2):838–844. doi: 10.1128/jb.114.2.838-844.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. KORN R. J., YAKULIS V. J., LEMKE C. E., CHOMET B. Cold agglutinins in Listeria monocytogenes infections. AMA Arch Intern Med. 1957 Apr;99(4):573–580. doi: 10.1001/archinte.1957.00260040073008. [DOI] [PubMed] [Google Scholar]
  15. Lind K. Production of cold agglutinins in rabbits induced by Mycoplasma pneumoniae, Listeria monocytogenes or Streptococcus MG. Acta Pathol Microbiol Scand B Microbiol Immunol. 1973 Aug;81(4):487–496. doi: 10.1111/j.1699-0463.1973.tb02234.x. [DOI] [PubMed] [Google Scholar]
  16. Lopes J., Inniss W. E. Electron microscopic study of lipopolysaccharide from an avian strain of Escherichia coli O18. J Bacteriol. 1970 Jul;103(1):238–243. doi: 10.1128/jb.103.1.238-243.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. MCILWAIN P., EVELETH D. F., DOUBLY J. A. PHARMACOLOGIC STUDIES OF A TOXIC CELLULAR COMPONENT OF LISTERIA MONOCYTOGENES. Am J Vet Res. 1964 May;25:774–781. [PubMed] [Google Scholar]
  18. Milner K. C., Finkelstein R. A. Bioassay of endotoxin: correlation between pyrogenicity for rabbits and lethality for chick embryos. J Infect Dis. 1966 Dec;116(5):529–536. doi: 10.1093/infdis/116.5.529. [DOI] [PubMed] [Google Scholar]
  19. Mára M., Julăk J., Patocka F. Isolation and chemical properties of biologically active fractions of the surface of Listeria monocytogenes. J Hyg Epidemiol Microbiol Immunol. 1974;18(3):359–364. [PubMed] [Google Scholar]
  20. Mára M., Patocka F., Julák J., Potuzníková B. Contribution to knowledge of factors participating in virulence of Listeria monocytogenes. II. Study of the chemical nature of complex Ei. J Hyg Epidemiol Microbiol Immunol. 1974;18(1):29–41. [PubMed] [Google Scholar]
  21. Olins A. L., Warner R. C. Physicochemical studies on a lipopolysaccharide from the cell wall of Azotobacter vinelandii. J Biol Chem. 1967 Nov 10;242(21):4994–5001. [PubMed] [Google Scholar]
  22. Owen P., Salton M. R. Isolation and characterization of a mannan from mesosomal membrane vesicles of Micrococcus lysodeikticus. Biochim Biophys Acta. 1975 Oct 6;406(2):214–234. doi: 10.1016/0005-2736(75)90006-1. [DOI] [PubMed] [Google Scholar]
  23. Patocka F., Mára M., Potuźníková B., Jirásek A., Mencíková E. Biological properties of surface components of Listeria monocytogenes (Ei factor). J Hyg Epidemiol Microbiol Immunol. 1974;18(3):365–368. [PubMed] [Google Scholar]
  24. Roberts R. S. Preparation of endotoxin. Nature. 1966 Jan 1;209(5018):80–80. doi: 10.1038/209080a0. [DOI] [PubMed] [Google Scholar]
  25. Shands J. W., Jr, Graham J. A., Nath K. The morphologic structure of isolated bacterial lipopolysaccharide. J Mol Biol. 1967 Apr 14;25(1):15–21. doi: 10.1016/0022-2836(67)90275-6. [DOI] [PubMed] [Google Scholar]
  26. Siddique I. H., Srivastava K. K. Cell wall and cytoplasmic components of Listeria monocytogenes. Am J Vet Res. 1973 Apr;34(4):567–570. [PubMed] [Google Scholar]
  27. Siddique I. H., Ying L. C., Chung R. A. Studies on diphosphopyridine nucleotidase and platelet damaging factor in an extracellular product of Listeria monocytogenes. Can J Microbiol. 1970 Oct;16(10):909–916. doi: 10.1139/m70-157. [DOI] [PubMed] [Google Scholar]
  28. Steck T. L., Yu J. Selective solubilization of proteins from red blood cell membranes by protein perturbants. J Supramol Struct. 1973;1(3):220–232. doi: 10.1002/jss.400010307. [DOI] [PubMed] [Google Scholar]
  29. Tsai C. M., Zopf D. A., Wistar R., Jr, Ginsburg V. A human cold agglutinin which binds lacto-N-neotetraose. J Immunol. 1976 Sep;117(3):717–721. [PubMed] [Google Scholar]
  30. Weckesser J., Drews G., Ladwig R. Localization and biological and physicochemical properties of the cell wall lipopolysaccharide of Rhodopseudomonas capsulata. J Bacteriol. 1972 Apr;110(1):346–353. doi: 10.1128/jb.110.1.346-353.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wicken A. J., Knox K. W. Lipoteichoic acids: a new class of bacterial antigen. Science. 1975 Mar 28;187(4182):1161–1167. doi: 10.1126/science.46620. [DOI] [PubMed] [Google Scholar]

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

RESOURCES