Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1991 Sep;59(9):2880–2884. doi: 10.1128/iai.59.9.2880-2884.1991

Mouse liver contains a Pseudomonas aeruginosa exotoxin A-binding protein.

J J Forristal 1, M R Thompson 1, R E Morris 1, C B Saelinger 1
PMCID: PMC258108  PMID: 1879914

Abstract

The opportunistic pathogen Pseudomonas aeruginosa produces several potential virulence factors, including the ADP-ribosylating toxin, exotoxin A (PE). Studies using a burned mouse model have shown that PE consistently inhibits protein synthesis and depletes elongation factor 2 in mouse liver and variably in other organs. One reason for toxin sensitivity could be the presence of a PE receptor on the surface of cells. Therefore we examined detergent extracts of mouse tissues for the presence of toxin-binding proteins. Proteins which specifically bind PE were present in extracts from liver, kidney, lung, spleen, and heart. Because liver appears to be a prominent target for the toxin in a burned animal, we choose to isolate the PE-binding protein from mouse liver and compare this protein to the recently characterized toxin-binding protein from toxin-sensitive mouse LM fibroblasts. The toxin-binding proteins from both sources have a molecular mass of approximately 350 kDa, share similar protease digestion profiles, and are glycosylated. However the glycosylation patterns for the two species are quite different. Both glycoproteins bind toxin with high avidity. The toxin-binding moiety is located, at least in part, on the plasma membrane and thus could represent the receptor involved in internalization of toxin molecules responsible for cell death.

Full text

PDF

Images in this article

2882Image
on p.2882
2882Image
on p.2882
2882Image
on p.2882
2883Image
on p.2883
2883Image
on p.2883

Selected References

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

  1. Croze E. M., Morré D. J. Isolation of plasma membrane, golgi apparatus, and endoplasmic reticulum fractions from single homogenates of mouse liver. J Cell Physiol. 1984 Apr;119(1):46–57. doi: 10.1002/jcp.1041190109. [DOI] [PubMed] [Google Scholar]
  2. FitzGerald D., Morris R. E., Saelinger C. B. Receptor-mediated internalization of Pseudomonas toxin by mouse fibroblasts. Cell. 1980 Oct;21(3):867–873. doi: 10.1016/0092-8674(80)90450-x. [DOI] [PubMed] [Google Scholar]
  3. Iglewski B. H., Kabat D. NAD-dependent inhibition of protein synthesis by Pseudomonas aeruginosa toxin,. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2284–2288. doi: 10.1073/pnas.72.6.2284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Iglewski B. H., Liu P. V., Kabat D. Mechanism of action of Pseudomonas aeruginosa exotoxin Aiadenosine diphosphate-ribosylation of mammalian elongation factor 2 in vitro and in vivo. Infect Immun. 1977 Jan;15(1):138–144. doi: 10.1128/iai.15.1.138-144.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  6. Marynen P., Van Leuven F., Cassiman J. J., Van den Berghe H. Solubilization and affinity purification of the alpha 2-macroglobulin receptor from human fibroblasts. J Biol Chem. 1984 Jun 10;259(11):7075–7079. [PubMed] [Google Scholar]
  7. Morris R. E., Saelinger C. B. Reduced temperature alters Pseudomonas exotoxin A entry into the mouse LM cell. Infect Immun. 1986 May;52(2):445–453. doi: 10.1128/iai.52.2.445-453.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Nørby J. G. Coupled assay of Na+,K+-ATPase activity. Methods Enzymol. 1988;156:116–119. doi: 10.1016/0076-6879(88)56014-7. [DOI] [PubMed] [Google Scholar]
  9. Ogata M., Chaudhary V. K., Pastan I., FitzGerald D. J. Processing of Pseudomonas exotoxin by a cellular protease results in the generation of a 37,000-Da toxin fragment that is translocated to the cytosol. J Biol Chem. 1990 Nov 25;265(33):20678–20685. [PubMed] [Google Scholar]
  10. Pavlovskis O. R., Pollack M., Callahan L. T., 3rd, Iglewski B. H. Passive protection by antitoxin in experimental Pseudomonas aeruginosa burn infections. Infect Immun. 1977 Dec;18(3):596–602. doi: 10.1128/iai.18.3.596-602.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Pavlovskis O. R., Shackelford A. H. Pseudomonas aeruginosa exotoxin in mice: localization and effect on protein synthesis. Infect Immun. 1974 Mar;9(3):540–546. doi: 10.1128/iai.9.3.540-546.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Pavlovskis O. R., Voelker F. A., Shackelford A. H. Pseudomonas aeruginosa exotoxin in mice: histopathology and serum enzyme changes. J Infect Dis. 1976 Mar;133(3):253–259. doi: 10.1093/infdis/133.3.253. [DOI] [PubMed] [Google Scholar]
  13. Pirker R., FitzGerald D. J., Hamilton T. C., Ozols R. F., Willingham M. C., Pastan I. Anti-transferrin receptor antibody linked to Pseudomonas exotoxin as a model immunotoxin in human ovarian carcinoma cell lines. Cancer Res. 1985 Feb;45(2):751–757. [PubMed] [Google Scholar]
  14. Rome L. H., Garvin A. J., Allietta M. M., Neufeld E. F. Two species of lysosomal organelles in cultured human fibroblasts. Cell. 1979 May;17(1):143–153. doi: 10.1016/0092-8674(79)90302-7. [DOI] [PubMed] [Google Scholar]
  15. Saelinger C. B., Snell K., Holder I. A. Experimental studies on the pathogenesis of infections due to Pseudomonas aeruginosa: direct evidence for toxin production during Pseudomonas infection of burned skin tissues. J Infect Dis. 1977 Oct;136(4):555–561. doi: 10.1093/infdis/136.4.555. [DOI] [PubMed] [Google Scholar]
  16. Snell K., Holder I. A., Leppla S. A., Saelinger C. B. Role of exotoxin and protease as possible virulence factors in experimental infections with Pseudomonas aeruginosa. Infect Immun. 1978 Mar;19(3):839–845. doi: 10.1128/iai.19.3.839-845.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Takatsuki A., Tamura G. Effect of tunicamycin on the synthesis of macromolecules in cultures of chick embryo fibroblasts infected with Newcastle disease virus. J Antibiot (Tokyo) 1971 Nov;24(11):785–794. doi: 10.7164/antibiotics.24.785. [DOI] [PubMed] [Google Scholar]
  18. Thompson M. R., Forristal J., Kauffmann P., Madden T., Kozak K., Morris R. E., Saelinger C. B. Isolation and characterization of Pseudomonas aeruginosa exotoxin A binding glycoprotein from mouse LM cells. J Biol Chem. 1991 Feb 5;266(4):2390–2396. [PubMed] [Google Scholar]
  19. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Vasil M. L., Iglewski B. H. Comparative toxicities of diphtherial toxin and Pseudomonas aeruginosa exotoxin A: evidence for different cell receptors. J Gen Microbiol. 1978 Oct;108(2):333–337. doi: 10.1099/00221287-108-2-333. [DOI] [PubMed] [Google Scholar]

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

RESOURCES