1887

Abstract

A study was conducted into whether or not nicking of the A subunit of LT enterotoxin at position Arg192 or its neighbouring amino acids Arg192 to The 195 is required for its toxicity. The toxic activity of mutants created by substitution or deletion at this position, which lacked ADP-ribosyltransferase activity was not completely obliterated and cyclic AMP was partially induced in the target cells, showing that they still displayed enzymic activity Moreover, although the A subunit possesses three potential sites for cleavage by furin, furin was not involved in the partial toxicity and cyclic AMP induction observed. These data suggest that target cells have a nick mechanism that operates at sites other than those around Arg192 or those recognized by furin, which generates an active fragment by processing the A subunit after toxin binding to the cell membrane.

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1997-06-01
2021-10-28
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References

  1. Booth, B. A.,, Boesman-Finkelstein, M., Finkelstein, R. A. 1984; Vibrio cholerae hemagglutinin/protease nicks cholera toxin.. Infect Immun 45:558–560
    [Google Scholar]
  2. Clements, J. D., Finkelstein, R. A. 1979; Isolation and characterization of homologous heat-labile enterotoxins with high specific activity from Escherichia coli cultures.. Infect Immun 24:760–769
    [Google Scholar]
  3. Craig, J. P. 1979; A permeability factor (toxin) found in cholera stools and culture filtrates and its neutralization by convalescent cholera sera.. Nature 207:614–616
    [Google Scholar]
  4. Dallas, W. S.,, Gill, D. M., Falkow, S. 1979; A permeability factor (toxin) found in cholera stools and culture filtrates and its neutralization by convalescent cholera sera.. Nature 207:614–616
    [Google Scholar]
  5. Dickinson, B., Clements, J. D. 1995; Dissociation of Escherichia coli heat-labile enterotoxin adjuvanticity from ADP-ribosyltransferase activity.. Infect Immun 63:1617
    [Google Scholar]
  6. Eagle, H. 1959; Amino acid metabolism in mammalian cell cultures.. Science 13:432–437
    [Google Scholar]
  7. Finkelstein, R. A. 1973; Cholera. Crit Rev Microbiol 2:553–623
    [Google Scholar]
  8. Finkelstein, R. A., Boesman-Finkelstein, M., Holt, P. 1983; Vibrio cholerae hemagglutinin/lectin/protease hydrolyzes fibronectin and ovomucin: F. M. Burnet revisited.. Proc Natl Acad Sci USA 80:1092–1095
    [Google Scholar]
  9. Freissmuth, M., Gilman, A. G. 1989; Mutations of G designed to alter the reactivity of the protein with bacterial toxins.. J Biol Chem 264:21907–21914
    [Google Scholar]
  10. Garred, O.,, van Deurs, B., Sandvig, K. 1995; Furin-induced cleavage and activation of shiga toxin.. J Biol Chem 270:10817–10821
    [Google Scholar]
  11. Gill, D. M., King, C.A. 1975; The mechanism of action of cholera toxin in pigeon erythrocyte lysates.. J Biol Chem 250:6424–6432
    [Google Scholar]
  12. Grant, C. C. R.,, Messer R.J., Cieplak, W. 1994; Role of trypsinlike cleavage at arginine 192 in the enzymatic and cytotonic activities of Escherichia coli heat-labile enterotoxin.. Infect Immun 62:4270–4278
    [Google Scholar]
  13. Ham, R. G. 1965; Clonal growth of mammalian cells in a chemically defined, synthetic medium.. Proc Natl Acad Sci USA 53:288–293
    [Google Scholar]
  14. Hirst, T. R.,, Randall, L. L., Hardy, S.J. 1984; Cellular location of heat-labile enterotoxin in Escherichia coli.. J Bacteriol 157:637–643
    [Google Scholar]
  15. Holmgren, J. 1981; Actions of cholera toxin and the prevention and treatment of cholera.. Nature 292:413–417
    [Google Scholar]
  16. Ichinose, Y.,, Tsuji, T.,, Ehara, M.,, Miyama, A., Naito, T. 1992; A protease from Vibrio cholerae nicks arginine at position 192 from the N-terminus of the heat-labile enterotoxin A subunit from enterotoxigenic Escherichia coli.. Eur J Epidemiol 8:743–747
    [Google Scholar]
  17. Ito, W.,, Ishiguro H., Kurosawa, Y. 1991; A general method for introducing a series of mutations into cloned DNA using the polymerase chain reaction.. Gene 102:67–70
    [Google Scholar]
  18. Klapper, D. G.,, Finkelstein, R. A., Capra, J. D. 1976; Subunit structure and N-terminal amino acid sequence of the three chains of cholera enterotoxin.. Immunochemistry 13:605–611
    [Google Scholar]
  19. Kunkel, S. L, Robertson, D. C. 1979; Purification and chemical characterization of the heat-labile enterotoxin produced by enterotoxigenic Escherichia coli.. Infect Immun 25:586–596
    [Google Scholar]
  20. Laemmli, U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature 227:680–685
    [Google Scholar]
  21. Lycke, N.,, Tsuji, T., Holmgren J. 1992; The adjuvant effect of Vibrio cholerae and Escherichia coli heat-labile enterotoxins is linked to their ADP-ribosyltransferase activity.. Eur J Immunol 22:2277–2281
    [Google Scholar]
  22. Mekalanos, J. J.,, Collier, R. J., Romig, W. R. 1979; Enzymatic activity of cholera toxin.. J Biol Chem 254:5855–5861
    [Google Scholar]
  23. Mekalanos, J. J.,, Swartz, D. J.,, Pearson, G. D. N.,, Harford, N.,, Groyne, F., de Wilde, M. 1983; Cholera toxin genes; nucleotide sequence, deletion analysis and vaccine development.. Nature 306:551–557
    [Google Scholar]
  24. Moss, J., Vaughan, M. 1990; ADP-ribosylating toxins and G proteins. Washington, DC : American Society for Microbiology..
    [Google Scholar]
  25. Moss, J., Garrison, S.,, Oppenheimer, N. J., Richardson, S. H. 1979; NAD-dependent ADP-ribosylation of arginine and proteins by Escherichia coli heat-labile enterotoxin.. J Biol Chem 254:6270–6272
    [Google Scholar]
  26. Moss, J., Stanley, S.J., Vaughan, M., Tsuji, T. 1993; Interaction of ADP-ribosylation factor with Escherichia coli enterotoxin that contains an inactivating lysine 112 substitution.. J Biol Chem 268:6383–6387
    [Google Scholar]
  27. Nakayama, K.,, Watanabe, T.,, Nakagawa, T.,, Kim, W. S.,, Nagahama, M.,, Hosaka, M.,, Hatsuzawa, K.,, Kondo-Hashiba, K., Murakami, K. 1992; Consensus sequence for precursor processing at mono-arginyl sites.. J Biol Chem 267:16335–16340
    [Google Scholar]
  28. Neill, R. J.,, Ivins, B. E., Holmes, R. K. 1983; Synthesis and secretion of the plasmid-coded heat-labile enterotoxin of Escherichia coli in Vibrio cholerae.. Science 221:289–290
    [Google Scholar]
  29. Orlandi, P. A.,, Curran, P. K., Fishman, P. H. 1993; Brefeldin A blocks the response of cultured cells to cholera toxin.. J Biol Chem 268:12010–12016
    [Google Scholar]
  30. Pearson, G. D. N., Mekalanos, J. J. 1982; Molecular cloning of Vibrio cholerae enterotoxin genes in Escherichia coli K-12.. Proc Natl Acad Sci USA 79:2976–2980
    [Google Scholar]
  31. Rappaport, R. S.,, Sagin, J. F.,, Pierzchala, W. A.,, Bonde, G.,, Rubin, B. A., Tint, H. 1976; Activation of heat-labile Escherichia coli enterotoxin by trypsin.. J Infect Dis 133:S41–S54
    [Google Scholar]
  32. Saiki, R. K.,, Scharf, S.,, Faloona, F.,, Mullis, K. B.,, Horn, G. T.,, Erlich, H.A., Arnheim, N. 1985; Enzymatic amplification of β-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia.. Science 230:1350–1354
    [Google Scholar]
  33. Sixma, T. K.,, Pronk, S. E.,, Kalk, K. H.,, Wartna, E. S.,, van Zanten, B. A. M.,, Witholt, B., Hol, W. C. J. 1991; Crystal structure of a cholera toxin-related heat-labile enterotoxin from Escherichia coli.. Nature 351:371–377
    [Google Scholar]
  34. Spangler, B. D. 1992; Structure and function of cholera toxin and the related Escherichia coli heat-labile enterotoxin.. Microbiol Rev 56:622–647
    [Google Scholar]
  35. Takeda, Y.,, Honda, T.,, Taga S., Miwatani, T. 1981; In vitro formation of hybrid toxins between subunits of Escherichia coli heat-labile enterotoxin and those of cholera toxin.. Infect Immun 34:341–346
    [Google Scholar]
  36. Tsuji, T.,, Honda, T.,, Miwatani, T. 1984; Comparison of effects of nicked and unnicked Escherichia coli heat-labile enterotoxin on Chinese hamster ovary cells.. Infect Immun 46:94–97
    [Google Scholar]
  37. Tsuji, T.,, Inoue, T.,, Miyama, A.,, Okamoto, K.,, Honda, T.,, Miwatani, T. 1990; A single amino acid substitution in the A subunit of Escherichia coli enterotoxin results in a loss of its toxic activity.. J Biol Chem 265:22520–22525
    [Google Scholar]
  38. Tsuji, T.,, Inoue, T.,, Miyama, A.,, Noda, M. 1991; Glutamic acid- 112 of the A subunit of heat-labile enterotoxin from enterotoxigenic Eschericha coli is important for ADP-ribosyltransferase activity.. FEBS Lett 291:319–321
    [Google Scholar]
  39. Tsuji, T.,, Kato, M.,, Kato, Y.,, Kawase, H.,, Imamura, S., Miyama, A. 1994; Construction of plasmids useful to the B subunit of cholera toxin from Vibrio cholerae or a heat-labile enterotoxin from enterotoxigenic Escherichia coli . Eur J Epidemiol 10:393–398
    [Google Scholar]
  40. Tsuneoka, M.,, Nakayama, K.,, Hatsuzawa, K.,, Komada, M.,, Kitamura, N., Mekada, E. 1993; Evidence for involvement of furin in cleavage and activation of diphtheria toxin.. J Biol Chem 268:26461–26465
    [Google Scholar]
  41. Uesaka, Y.,, Otsuka, T.,, Lin, Z.,, Yamazaki, S.,, Yamaoka, J.,, Kurazono, H., Takeda, Y. 1994; Simple method of purification of Escherichia coli heat-labile enterotoxin and cholera toxin using immobilized galactose.. Microb Pathog 16:71–76
    [Google Scholar]
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