Chapter 23 : Holistic Perspective on the Escherichia coli Hemolysin

MyBook is a cheap paperback edition of the original book and will be sold at uniform, low price.

Preview this chapter:
Zoom in

Holistic Perspective on the Escherichia coli Hemolysin, Page 1 of 2

| /docserver/preview/fulltext/10.1128/9781555818340/9781555810825_Chap23-1.gif /docserver/preview/fulltext/10.1128/9781555818340/9781555810825_Chap23-2.gif


This chapter reviews the knowledge of the prototype of the RTX family of pore-forming toxins, the hemolysin, and proposes that new efforts be made to study the significance and roles of the exotoxins, together with those of endotoxins, as an integrated synergistic system of multiple virulence factors. The fundamental approach taken by biochemists and cell biologists interested in understanding the role in pathogenesis of a bacterial virulence factor such as an exoprotein has been to purify the exoprotein and observe its activity on different substrates, cells, and hosts. The RTX-associated diseases include pertussis, juvenile periodontitis, pneumonia, urinary tract infections, and wound infections. The epidemiological evidence for an association between RTX toxin production and isolates that cause disease is strong for all of the organisms except i, , and . The solubilization of hemolysin aggregates by a chaotropic agent such as urea causes an increase in the specific lytic activity of the hemolysin. The erythrocytes transform into echinocytes, in which gross cytoskeletal or membrane irregularities result in rounding of the cell and the formation of multiple teardrop-shaped projections from the cell surface. The cellular transformation is similar to that caused by the treatment of erthrocytes by a calcium ionophore. The motivation for covering the older literature in the chapter is to stimulate thought, experimentation, and discussion about the significance of the ever-present lipopolysaccharide (LPS) molecules on the structure and activity of the hemolysin.

Citation: Welch R. 1994. Holistic Perspective on the Escherichia coli Hemolysin, p 351-364. In Miller V, Kaper J, Portnoy D, Isberg R (ed), Molecular Genetics of Bacterial Pathogenesis. ASM Press, Washington, DC. doi: 10.1128/9781555818340.ch23
Highlighted Text: Show | Hide
Loading full text...

Full text loading...


Image of Figure 1
Figure 1

After a 5-min treatment with the hemolysin, sheep erythrocytes were fixed and visualized by scanning electron microscopy ( ). The photograph shows the loss of the concave cellular structure typical of erythrocytes and the formation of multiple teardrop-shaped projections from the cell surface. Bar, 2 μm. The photograph was the gift of Sally Jorgensen, University of Minnesota.

Citation: Welch R. 1994. Holistic Perspective on the Escherichia coli Hemolysin, p 351-364. In Miller V, Kaper J, Portnoy D, Isberg R (ed), Molecular Genetics of Bacterial Pathogenesis. ASM Press, Washington, DC. doi: 10.1128/9781555818340.ch23
Permissions and Reprints Request Permissions
Download as Powerpoint


1. Arthur, M.,, C. E. Johnson,, R. H. Rubin,, R. D. Arbeit,, C. Campanelli,, C. Kim,, S. Steinbach,, M. Agarwal,, R. Wilkinson,, and R. Goldstein. 1989. Molecular epidemiology of adhesin and hemolysin virulence factors among uropathogenic Escherichia coli. Infect. Immun. 57: 303 313.
2. Benz, R.,, A. Schmid,, W. Wagner,, and W. Goebel. 1989. Pore formation by the Escherichia coli hemolysin: evidence for an association-dissociation equilibrium of the pore-forming aggregates. Infect. Immun. 57: 887 895.
3. Bhakdi, S. Unpublished data.
4. Bhakdi, S.,, N. Mackman,, J. M. Nicaud,, and I. B. Holland. 1986. Escherichia coli hemolysin may damage target cell membranes by generating transmembrane pores. Infect. Immun. 52: 63 69.
5. Bhakdi, S.,, and E. Martin. 1991. Superoxide generation by human neutrophils induced by low doses of Escherichia coli hemolysin. Infect. Immun. 59: 2955 2962.
6. Bohach, G. A.,, and I. S. Snyder. 1985. Chemical and immunological analysis of thecomplex structure of Escherichia coli a-hemolysin. J. Bacteriol. 164: 1071 1080.
7. Bohach, G.,, and I. S. Snyder. 1986. Composition of affinity-purified a-hemolysin of Escherichia coli. Infect. Immun. 53: 435 437.
8. Burrows, L. L.,, and R. Y. C. Lo. 1992. Molecular characterization of an RTX toxin determinant from Actinobacillus suis. Infect. Immun. 60: 2166 2173.
9. Camprubi, S.,, J. Tomas,, F. Munoa,, C. Madrid,, and A. Juarez. 1990. Influence of lipopolysaccha-ride on external hemolytic activity of Salmonella typhimurium and Klebsiella pneumoniae. Curr. Microbiol. 20: 1 3.
10. Cavalier!, S. J.,, and I. S. Snyder. 1982. Cytotoxic activity of partially purified Escherichia coli alpha haemolysin. J. Med. Microbiol. 15: 11 21.
11. Chang, Y.,, D. Ma,, J. Shi,, and M. M. Chengappa. 1993. Molecular characterization of a leukotoxin gene from a Pasteurella haemolytica-like organism, encoding a new member of the RTX toxin family. Infect. Immun. 61: 2089 2095.
12. Chang, Y.-F.,, R. Young,, D. Post,, and D. K. Struck. 1987. Identification and characterization of the Pasteurella haemolytica leukotoxin. Infect. Immun. 55: 2348 2354.
13. Chang, Y.-F.,, R. Young,, and D. K. Struck. 1989. Cloning and characterization of a hemolysin gene from Actinobacillus (Haemophilus) pleuropneumoniae. DNA 8: 635 647.
14. Falkow, S. 1988. Molecular Koch's postulates applied to microbial pathogenicity. Rev. Infect. Dis. 10: S274 S276.
15. Fasano, A.,, B. Baudry,, D. Pumplin,, S. Wasserman,, B. D. Tall,, J. Ketley,, and J. B. Kaper. 1991. Vibrio cholerae produces a second enterotoxin, which affects intestinal tight junctions. Proc. Natl. Acad. Sci. USA 88: 5242 5246.
16. Felmlee, T.,, S. Pellett,, E. Y. Lee,, and R. A. Welch. 1985. The Escherichia coli hemolysin is released extracellularly without cleavage of a signal peptide. J. Bacteriol. 163: 88 93.
17. Felmlee, T.,, S. Pellett,, and R. A. Welch. 1985. The nucleotide sequence of an Escherichia coli chromosomal hemolysin. J. Bacteriol. 163: 94 105.
18. Fontaine, A.,, J. Arondel,, and P. J. Sansonetti. 1988. Role of Shiga toxin in the pathogenesis of bacillary dysentery, studied by using a Tox - mutant of Shigella dysenteriae 1. Infect. Immun. 56: 3099 3109.
19. Frey, J.,, R. Meier,, D. Gygi,, and J. Nicolet. 1991. Nucleotide sequence of the hemolysin I gene from Actinobacillus pleuropneumoniae. Infect. Immun. 59: 3026 3032.
20. Glaser, P.,, D. Ladant,, O. Sezer,, F. Pichot,, A. Ullman,, and A. Danchin. 1988. The calmodulin-sensitive adenylate cyclase of Bordetella pertussis: cloning and expression in Escherichia coli. Mol. Microbiol. 2: 19 30.
21. Gonzalez-Carrero, M. I.,, J. Zabala,, F. de la Cruz,, and J. M. Ortiz. 1985. Purification of a hemolysin from an overproducing E. coli strain. Mol. Gen. Genet. 109: 106 110.
22. Grlmminger, F.,, U. Sibelius,, S. Bhakdi,, N. Suttorp,, and W. Seeger. 1991. Escherichia coli hemolysin is a potent inductor of phosphoinositide hydrolysis and related metabolic responses in human neutrophils. J. Clin. Invest. 88: 1531 1539.
23. Hacker, J. C.,, C. Hughes,, H. Hof,, and W. Goebel. 1983. Cloned hemolysin genes from Escherichia coli that cause urinary tract infection determine different levels of toxicity in mice. Infect. Immun. 42: 57 63.
24. Hewlett, E. L.,, V. M. Gordon,, J. D. McCaffery,, W. M. Sutherland,, and M. C. Gray. 1989. Adenylate cyclase toxin from Bordetella pertussis: identification of the holotoxin molecule. J. Biol. Chem. 264: 19379 19384.
25. Highlander, S. K.,, M. Chidambaram,, M. J. Engler,, and G. M. Weinstock. 1989. DNA sequence of the Pasteurella haemolytica leukotoxin gene cluster. DNA 8: 15 28.
26. Holmgren, J.,, and A. Svennerholm. 1977. Mechanism of disease and immunity in cholera: a review. J. Infect. Dis. 136S: 105 112.
27. Issartel, J.-P.,, V. Koronakis,, and C. Hughes. 1991. Activation of Escherichia coli prohaemolysin to the mature toxin by acyl carrier protein-dependent fatty acylation. Nature (London) 351: 759 761.
28. Jansen, R.,, J. Brake,, E. M. Kamp,, A. L. Gielkins,, and M. A. Smits. 1993. Cloning and characterization of the Actinobacillus pleuropnettmoniae RTX-toxin III (ApxIII) gene. Infect. Immun. 61: 947 954.
29. Jonas, D.,, B. Schultheis,, C. Klas,, P. H. Krammer,, and S. Bhakdi. 1993. Cytocidal effects of Escherichia coli hemolysin on human T lymphocytes. Infect. Immun. 61: 1715 1721.
30. Jorgensen, S. E.,, R. F. Hammer,, and G. K. Wu. 1980. Effects of a single hit from the α-hemolysin produced by Escherichia coli on the morphology of sheep erythrocytes. Infect. Immun. 27: 988 994.
31. Jorgensen, S. E.,, P. F. Muicahy,, G. K. Wu,, and C. F. Louis. 1983. Calcium accumulation in human and sheep erythrocytes that is induced by Escherichia coli hemolysin. Toxicon 21: 717 727.
32. Kaper, J. B.,, H. Lockman,, M. M. Baldini,, and M. M. Levine. 1984. A recombinant live oral cholera vaccine. Bio/Technology 2: 345 349.
33. Kaper, J. B.,, H. Lockman,, M. M. Baldini,, and M. M. Levine. 1984. Recombinant nontoxigenic Vibrio cholerae strains as attenuated cholera vaccines candidates. Nature (London) 308: 655 658.
34. Koronakis, V.,, M. Cross,, B. Senior,, E. Koranakis,, and C. Hughes. 1987. The secreted hemolysins of Proteus mirabilis, Proteus vulgaris, and Morganella morganii are genetically related to each other and to the α-hemolysin of Escherichia coli. J. Bacteriol. 169: 1509 1515.
35. Kraig, E.,, T. Dailey,, and D. Kolodrubetz. 1990. Nucleotide sequence of the leukotoxin gene from Actinobacillus actinomycetemcomitans: homology to the α-hemolysin/leukotoxin gene family. Infect. Immun. 58: 920 929.
36. Levine, M. M.,, J. B. Kaper,, D. Herrington,, G. Losonsky,, J. G. Morris,, M. L. Clements,, R. E. Black,, B. Tall,, and R. Hall. 1988. Volunteer studies of deletion mutants of Vibrio cholerae O1 prepared by recombinant techniques. Infect. Immun. 56: 161 167.
37. Lo, R. Y., C. C. Strathdee,, and P. Shewen. 1987. Nucleotide sequence of the leukotoxin genes of Pasteurella haemolytica Al. Infect. Immun. 55: 1987 1996.
38. Ludwig, A.,, R. Benz,, and W. Goebel. 1993. Oligomerization of Escherichia coli (HlyA) is involved in pore formation. Mol. Gen. Genet. 241: 89 96.
39. Menestrina, G.,, N. Mackman,, I. B. Holland,, and S. Bhakdi. 1987. Escherichia coli haemolysin forms voltage-dependent ion channels in lipid membranes. Biochim. Biophys. Acta 905: 109 117.
40. Moayeri, M. Unpublished data.
41. Nicaud, J. M.,, N. Mackman,, L. Gray,, and I. B. Holland. 1985. Characterization of HlyC and mechanism of activation and secretion of hemolysin from E. coli 2001. FEBS Lett. 187: 339 344.
42. Opal, S. M.,, A. S. Cross,, P. Gemski,, and L. W. Lyhte. 1990. Aerobactin and a-hemolysin as virulence determinants in Escherichia coli isolated from human blood, urine, and stool. J. Infect. Dis. 161: 794 796.
43. Ostolaza, H.,, B. Bartolome,, I. O. de Zarate,, F. de la Cruz,, and F. Goni. 1993. Release of lipid vesicle contents by the bacterial protein toxin a-hemolysin. Biochim. Biophys. Acta 1147: 81 88.
44. Ostolaza, H.,, B. Bartoleme,, J. Serra,, F. de la Cruz,, and F. Goni. 1991. a-Hemolysin from E. coli: purification and self-aggregation properties. FEBS Lett. 280: 195 198.
45. Parker, C. T.,, A. W. Kloser,, C. A. Schnaitman,, M. A. Stein,, S. Gottesman,, and B. W. Gibson. 1992. Role of the rfaG and rfaP genes in determination of the lipolysaccharide structure and cell surface properties of Escherichia coli. J. Bacteriol. 174: 2525 2538.
46. Rennie, R. P.,, and J. P. Arbuthnott. 1974. Partial characterization of Escherichia coli haemolysin. J. Med. Microbiol. 7: 179 188.
47. Rennie, R. P.,, J. H. Freer,, and J. P. Arbuthnott. 1974. The kinetics of erythrocyte lysis by Escherichia coli haemolysin. J. Med. Microbiol. 7: 189 195.
48. Rocque, W.,, R. Coughlin,, and E. McGroarty. 1987. Lipopolysaccharide tightly bound to porin monomers and trimers from Escherichia coli K-12. J. Bacteriol. 169: 4003 4010.
49. Rogel, A.,, J. Schultz,, R. M. Brownlie,, J. G. Coote,, R. Parton,, and E. Hanski. 1989. Bordetella pertussis adenylate cyclase: purification and characterization of the toxic form of the enzyme. EMBO J. 8: 2755 2760.
50. Short, E. C. Jr.,, and H. J. Kurtz. 1971. Properties of the hemolytic activities of Escherichia coli. Infect. Immun. 3: 678 687.
51. Smith, H. W.,, and M. B. Huggins. 1985. The toxic role of alpha-haemolysin in the pathogenesis of experimental Escherichia coli infection in mice. J. Gen. Microbiol. 131: 395 403.
52. Stanley, P. L. D.,, P. Diaz,, M. J. A. Bailey,, D. Gygi,, A. Juarez,, and C. Hughes. 1993. Loss of activity in the secreted form of Escherichia coli haemolysin caused by an rfaP lesion in core lipopolysaccharide assembly. Mol. Microbiol. 10: 781 787.
53. Swihart, K. G.,, and R. A. Welch. 1990. The HpmA hemolysin is more common than HlyA among Proteus isolates. Infect. Immun. 58: 1853 1860.
54. Tesh, V. L.,, and A. D. O'Brien. 1991. The pathogenic mechanisms of Shiga toxin and the Shiga-like toxins. Mol. Microbiol. 5: 1817 1822.
55. Trucksis, M.,, J. E. Galen,, J. Michalski,, A. Fasano,, and J. Kaper. 1993. Accessory cholera entero-toxin (Ace), the third toxin of a Vibrio cholerae virulence cassette. Proc. Natl. Acad. Sci. USA 90: 5267 5271.
56. Wagner, W.,, M. Kuhn,, and W. Goebel. 1988. Active and inactive forms of hemolysin (HlyA) from Escherichia coli. Biol. Chem. Hoppe-Seyler 369: 39 46.
57. Wandersman, C.,, and S. Letoffe. 1993. Involvement of lipopolysaccharide in the secretion of Escherichia coli a-hemolysin and Erwinia chrysanthemi protease. Mol. Microbiol. 7: 141 150.
58. Welch, R. A. 1987. Identification of two different hemolysin determinants in uropathogenic Proteus isolates. Infect. Immun. 55: 2183 2190.
59. Welch, R. A. 1991. Pore-forming cytolysins of gram-negative bacteria. Mol. Microbiol. 5: 521 528.
60. Welch, R. A.,, E. P. Dellinger,, B. Minshew,, and S. Falkow. 1981. Hemolysin contributes to virulence of extra-intestinal Escherichia coli infections. Nature (London) 294: 665 667.
61. Welch, R. A.,, R. Hull,, and S. Falkow. 1983. Molecular cloning and physical characterization of a chromosomal hemolysin from Escherichia coli. Infect. Immun. 42: 178 186.
62. Welch, R. A.,, S. Pellett,, D. Robbins,, W. Keane,, G. Gekker,, and P. Peterson,. 1989. Epidemiological observations involving the Escherichia coli hemolysin. In E. Kass, and C. Svanborg-Eden (ed.), Host-Parasite Interactions in Urinary Tract Infections. University of Chicago Press, Chicago.
63. White, J. G. 1974. Effects of an ionophore, A23187, on the surface morphology of normal erythrocytes. Am. J. Pathol. 77: 507 518.
64. Williams, P. H. 1979. Determination of the molecular weight of Escherichia coli α-haemolysin. FEMS Microbiol. Lett. 5: 21 24.
65. Zwadyk, P.,, and I. S. Snyder. 1971. Purification and kinetic studies of the hemolysin from Escherichia coli. Can. J. Microbiol. 17: 741 745.


Generic image for table
Table 1

Selected features of toxins from the RTX toxin branch of the RTX exoprotein family

The predicted pi, based on the predicted amino acid sequence of the toxin gene product.

PMNs, polymorphonuclear leukocytes.

A weak hemolytic activity is observed.

No hemolytic activity is observed.

Citation: Welch R. 1994. Holistic Perspective on the Escherichia coli Hemolysin, p 351-364. In Miller V, Kaper J, Portnoy D, Isberg R (ed), Molecular Genetics of Bacterial Pathogenesis. ASM Press, Washington, DC. doi: 10.1128/9781555818340.ch23

This is a required field
Please enter a valid email address
Please check the format of the address you have entered.
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error