1887

Abstract

Shigellosis is characterized by a strong inflammatory response which is induced by bacteria invading the colonic mucosa. Characterization of a sepA mutant indicated that SepA, the major protein secreted by Shigella flexneri growing in laboratory media, might be involved in invasion and destruction of the host intestinal epithelium. The sequence of the first 500 residues of mature SepA (110 kDa) is homologous to that of the N-terminal region of lgA1 proteases. To investigate the potential proteolytic activity of SepA, the activity of the purified protein on a wide range of synthetic peptides was tested. SepA hydrolysed several of these substrates and the activity was inhibited by PMSF. Several peptides which were hydrolysed by SepA have been described as specific substrates for cathepsin G, a serine protease produced by polymorphonuclear leukocytes that was proposed to play a role in inflammation. However, unlike cathepsin G, SepA degraded neither fibronectin nor angiotensin I and had no effect on aggregation of human platelets. In addition, analysis of SepA hydrolysis by proteinase K suggested that the protein is composed of two domains of about 450 residues separated by a hinge region of 100 residues. The 47 kDa N-terminal domain was stable and endowed with proteolytic activity.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-144-7-1815
1998-07-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/micro/144/7/mic-144-7-1815.html?itemId=/content/journal/micro/10.1099/00221287-144-7-1815&mimeType=html&fmt=ahah

References

  1. Bachovchin W.W., Plaut A.G., Flentke G.R., Lynch M., Kettner C.A. 1990; Inhibition of IgAl proteinases from Neisseria gonorrhoeae and Haemophilus influenzae by peptide prolyl boronic acids.. J Biol Chern 265:3738–3743
    [Google Scholar]
  2. Baugh R.J., Travis J. 1976; Human leukocyte granule elastase: rapid isolation and characterization.. Biochemistry 15:836–841
    [Google Scholar]
  3. Benjelloun-Touimi Z., Sansonetti P.J., Parsot C. 1995; SepA, the major extracellular protein of Shigella flexneri: autonomous secretion and involvement in tissue invasion.. Mol Microbiol 17:123–125
    [Google Scholar]
  4. Bernardini M.L, Mounier J., d’Hauteville H., Conquis-Rondon M., Sansonetti P.J. 1989; Identification of icsA, a plasmid locus of Shigella flexneri that governs bacterial intra and intercellular spread through interaction with F-actin.. Proc Natl Acad Sci USA 863867–3871
    [Google Scholar]
  5. Bradford M.M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.. Anal Biochem 72:248–254
    [Google Scholar]
  6. Brunder W., Schmidt H., Karch H. 1997; EspP, a novel extracellular serine protease of enterohaemorrhagic Escherchia coli O157:H7 cleaves human coagulation factor V.. Mol Microbiol 24:767–778
    [Google Scholar]
  7. Djafari S., Ebel F., Deibel C., Krämer S., Hudel M., Chakraborty T. 1997; Characterization of an exported protease from Shiga toxin-producing Escherichia coli. . Mol Microbiol 25:771–784
    [Google Scholar]
  8. Erlanger B.F., Kokowsky N., Cohen W. 1961; The preparation and properties of two new chromogenic substrates of trypsin.. Arch Biochem Biophys 95:271–278
    [Google Scholar]
  9. Kielty C.M., Lees M., Shuttleworth C.A., Woolley D. 1993; Catabolism of intact type IV collagen microfibrils: susceptibility to degradation by serine proteinases.. Biochem Biophys Res Commun 191:1230–1236
    [Google Scholar]
  10. Klickstein L.B., Kaempfer C.E., Wintroub B.U. 1982; The granulocyte-angiotensin system. Angiotensin I-converting activity of cathepsin G.. J Biol Chern 257:15042–15046
    [Google Scholar]
  11. LaBrec E.H., Schneider H., Magnani T.J., Formal S.B. 1964; Epithelial cell penetration as an essential step in the pathogenesis of bacillary dysentery.. J Bacterial 88:1503–1518
    [Google Scholar]
  12. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature 227:680–685
    [Google Scholar]
  13. Lett M.C., Sasakawa C., Kamata K., Kurata T., Yoshikawa M. 1989; virG, a plasmid-coded virulence gene of Shigella flexneri: identification of the VirG protein and determination of the complete coding sequence.. J Bacteriol 171:353–359
    [Google Scholar]
  14. Ménard R., Sansonetti P.J., Parsot C. 1993; Nonpolar mutagenesis of the ipa genes defines IpaB, IpaC, and IpaD as effectors of Shigella flexneri entry into epithelial cells.. J Bacteriol 175:5899–5906
    [Google Scholar]
  15. Ménard R., Sansonetti P.J., Parsot C. 1994; The secretion of the Shigella flexneri Ipa invasins is activated by epithelial cells and controlled by IpaB and IpaD.. EMBO J 13:5293–5302
    [Google Scholar]
  16. Ménard R., Prévost M., Gounon P., Sansonetti P.J., Dehio C. 1996; The secreted Ipa complex of Shigella flexneri promotes entry into mammalian cells.. Proc Natl Acad Sci USA 931254–1258
    [Google Scholar]
  17. Molino M., Blanchard N., Belmonte E., Tarver A.P., Abrams C., Hoxie J.A., Cerletti C., Brass L.F. 1995; Proteolysis of the human platelet and endothelial cell thrombin receptor by neutrophil-derived cathepsin G.. J Biol Chem 270:11168–11175
    [Google Scholar]
  18. Mounier J., Vasselon T., Hellio R., Lesourd M., Sansonetti P.J. 1992; Shigella flexneri enters human colonic epithelial cells through the basolateral pole.. Infect Immun 60:237–248
    [Google Scholar]
  19. Nakajima K., Powers J.C., Ashe B.M., Zimmerman M. 1979; Mapping the extended substrate binding site of cathepsin G and human leukocyte elastase.. J Biol Chem 254:4027–4032
    [Google Scholar]
  20. Parsot C., Sansonetti P.J. 1995; Invasion and the pathogenesis of Shigella infections.. Curr Top Microbiol Immunol 209:25–42
    [Google Scholar]
  21. Perdomo O. J. J., Cavaillon J. M., Huerre M., Ohayon H., Gounon P., Sansonetti P. J. 1994a; Acute inflammation causes epithelial invasion and mucosal destruction in experimental shigellosis.. J Exp Med 180:1307–1319
    [Google Scholar]
  22. Perdomo O. J. J., Gounon P., Sansonetti P. J. 1994b; Poly-morphonuclear leukocyte transmigration promotes invasion of colonic epithelial monolayer by Shigella flexneri. . J Clin Invest 93:633–643
    [Google Scholar]
  23. Pohlner J., Halter R., Beyreuther K., Meyer T.F. 1987; Gene structure and extracellular secretion of Neisseria gonorrhoeae IgA protease.. Nature 325:458–462
    [Google Scholar]
  24. Poulsen K., Brandt J., Hjorth J.P., Thogersen H.C., Kilian M. 1989; Cloning and sequencing of the IgAl protease gene (iga) of Haemophilus influenzae serotype b.. Infect Immun 51:3097–3105
    [Google Scholar]
  25. Provence D., Curtis R. 1994; Isolation and characterization of a gene involved in hemagglutination by an avian pathogenic Escherichia coli strain.. Infect Immun 62:1369–1380
    [Google Scholar]
  26. Sansonetti P.J., Arondel J., Fontaine A., d’Hauteville H., Bernardini M.L. 1991; ompB (osmo-regulation) and icsA (cell- to-cell spread) mutants of Shigella flexneri: vaccine candidates and probes to study the pathogenesis of shigellosis.. Vaccine 9:416–422
    [Google Scholar]
  27. Selak M.A. 1994; Cathepsin G and thrombin: evidence for two different platelet receptors.. Biochem J 297:269–275
    [Google Scholar]
  28. Selak M.A., Chignard M., Smith J.B. 1988; Cathepsin G is a strong platelet agonist released by neutrophils.. Biochem J 251:293–299
    [Google Scholar]
  29. St Geme J.W. III de la Morena M.L., Falkow S. 1994; A Haemophilus influenzae IgA protease-like protein promotes intimate interaction with human epithelial cells.. Mol Microbiol 14:217–233
    [Google Scholar]
  30. Stein M., Kenny B., Stein M.A., Finlay B. 1996; Characterization of EspC, a 110-kilodalton protein secreted by enteropathogenic Escherichia coli which is homologous to members of the Immunoglobulin A protease-like family of secreted proteins.. J Bacteriol 178:6546–6554
    [Google Scholar]
  31. Tanaka T., Minematsu Y., Reilly C.F., Travis J., Powers J.C. 1985; Human leucocyte cathepsin G. Subsite mapping with 4- nitroanilides, chemical modification, and effect of possible cofactors.. Biochemistry 24:2040–2047
    [Google Scholar]
  32. Vartio T., Seppa H., Vaheri A. 1981; Susceptibility of soluble and matrix fibronectins to degradation by tissue proteinases, mast cell chymase and cathepsin G.. J Biol Chem 256:471–477
    [Google Scholar]
  33. Vu T.K.H., Hung D.T., Wheaton V.I., Coughlin S.R. 1991; Molecular cloning of a functional thrombin receptor reveals a novel proteolytic mechanism of receptor activation.. Cell 64:1057–1068
    [Google Scholar]
  34. Wassef J.S., Keren D.F., Mailloux J.L. 1989; Role of M cells in initial antigen uptake and in ulcer formation in the rabbit intestinal loop model of shigellosis.. Infect Immun 57:858–863
    [Google Scholar]
  35. Watarai M., Tobe T., Yoshikawa M., Sasakawa C. 1995; Contact of Shigella with host cells triggers release of Ipa invasins and is an essential function of invasiveness.. EMBO J 14:2461–2470
    [Google Scholar]
  36. Wood S.W., Burton J. 1991; Synthetic peptide substrates for the immunoglobulin Al protease from Neisseria gonorrhoeae (Type 2).. Infect Immun 59:1818–1822
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-144-7-1815
Loading
/content/journal/micro/10.1099/00221287-144-7-1815
Loading

Data & Media loading...

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