Skip to content
1887

Journal of Medical Microbiology logo Journal of Medical Microbiology

Volume 46, Issue 3

Isolation of a contact-dependent haemolysin from No Access

Abstract

Contact-dependent haemolytic activity was observed with cells of HRv and HRa, but not with those of BCG and . Culture filtrates of all these strains did not exhibit any haemolytic activity. HRv was subsequently used for the isolation of haemolysin. Haemolytic activity was retained in the cell debris even after sonication of the cells and treatment with Tween 80 and lysozyme. Solubilisation of haemolysin was possible only after the cell debris was washed with ethanol 70% and then treated with Tween 80 0.1%. The haemolysin thus obtained showed a micellar M of >200 000 by gelfiltration on Sephadex G-200 and a subunit M of 66 000 by SDS-PAGE. It was sensitive to trypsin but stable when heated at 60C for 10 min. Polyclonal serum raised in rabbits against the haemolysin neutralised the haemolytic activity. The N-terminal amino-acid sequence of the 66-kDa subunit of haemolysin showed identity with TB66, the 66-kDa secretory protein of , and 30% homology with the haemolysin A precursor of . Phosphatidylglycerol inhibited lysis of sheep erythrocytes by the haemolysin and is probably the receptor for the haemolysin. Haemolysin not only lysed erythrocytes, but was also cytotoxic to human lung cells. It appears that, among the members of the complex, the cell-bound contact-dependent haemolysin/cytolysin is restricted to and it may be associated with the pathogenesis of .

  • Received:
  • Accepted:
  • Published Online:
© 1997 The Pathological Society of Great Britain and Ireland
Loading

Article metrics loading...

/content/journal/jmm/10.1099/00222615-46-3-233
1997-03-01
2025-05-16
Loading full text...

Full text loading...

References

  1. Cossart P., Vicente M. F., Mengaud J., Baquero F., Perez-Diaz J. C., Berche P. Listeriolysin O is essential for virulence of Listeria monocytogenes: direct evidence obtained by gene complementation. Infect Immun 1989; 57:3629–3636
     [Google Scholar]
  2. Welch R. A. Pore-forming cytolysins of gram-negative bacteria. Mol Microbiol 1991; 5:521–528
     [Google Scholar]
  3. Sansonetti P. J., Ryter A., Clerc P., Maurelli A. T., Mounier J. Multiplication of Shigella flexneri with HeLa cells: lysis of the phagocytic vacuole and plasmid-mediated contact hemolysis. Infect Immun 1986; 51:461–469
     [Google Scholar]
  4. Welch R. A., Falkow S. Characterization of Escherichia coli hemolysins conferring quantitative differences in virulence. Infect Immun 1984; 43:156–160
     [Google Scholar]
  5. King C. H., Mundayoor S., Crawford J. T., Shinnick T. M. Expression of contact-dependent cytolytic activity by Mycobacterium tuberculosis and isolation of the genomic locus that encodes the activity. Infect Immun 1993; 61:2708–2712
     [Google Scholar]
  6. Udou T. Extracellular hemolytic activity in rapidly growing mycobacteria. Can J Microbiol 1994; 40:318–321
     [Google Scholar]
  7. Deshpande R. G., Khan M. B., Bhat D. A., Navalkar R. G. Purification and partial characterization of a novel 66-kDa seroreactive protein of Mycobacterium tuberculosis H37RV. J Med Microbiol 1994; 41:173–178
     [Google Scholar]
  8. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 1979; 76:4350–4354
     [Google Scholar]
  9. Howard S. P., Buckley J. T. Activation of the hole-forming toxin aerolysin by extracellular processing. J Bacteriol 1985; 163:336–340
     [Google Scholar]
  10. Braun V., Hobbie S., Ondraczek R. Serratia marcescens forms a new type of cytolysin. FEMS Microbiol Lett 1992; 100:299–306
     [Google Scholar]
  11. Harlow E., Lane D. Antibodies: a laboratory manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory; 1988100–102
     [Google Scholar]
  12. Leão S. C., Rocha C. L., Murillo L. A., Parra C. A., Patarroyo M. E. A species-specific nucleotide sequence of Mycobacterium tuberculosis encodes a protein that exhibits hemolytic activity when expressed in Escherichia coli . Infect Immun 1995; 63:4301–4306
     [Google Scholar]
  13. McDonough K. A., Kress Y. Cytotoxicity for lung epithelial cells is a virulence-associated phenotype of Mycobacterium tuberculosis . Infect Immun 1995; 63:4802–4811
     [Google Scholar]
  14. Mims C. A. The pathogenesis of infectious disease. 3rd edn London: Academic Press; 1990
     [Google Scholar]
/content/journal/jmm/10.1099/00222615-46-3-233
Loading
Isolation of a contact-dependent haemolysin from Mycobacterium tuberculosis
J. Med. Microbiol. 46, 233 (1997); https://doi.org/10.1099/00222615-46-3-233
/content/journal/jmm/10.1099/00222615-46-3-233
/content/journal/jmm/10.1099/00222615-46-3-233
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error