1887

Microbiology Society logo

Volume 148, Issue 2

A high-molecular-mass cell-surface protein from 1063 adheres to mucus components

The GenBank accession number for the sequence reported in this paper is AF120104.

Free

Abstract

A gene from 1063 encoding a cell-surface protein, designated Mub, that adheres to mucus components has been cloned and sequenced. The deduced amino acid sequence of Mub (358 kDa) shows the presence of 14 approximately 200 aa repeats and features typical for other cell-surface proteins of Gram-positive bacteria. Fusion proteins consisting of different repeats of Mub and the maltose-binding protein (MBP) were produced. These proteins adhered to pig mucus components, with molecular masses ranging from <01 to >2 MDa, to pig gastric mucin and to hen intestinal mucus. The binding of Mub to mucus components occurred in the pH range 3–74, with maximum binding at pH 4–5 and could be partly inhibited by the glycoprotein fetuin. Affinity-purified antibodies against recombinant Mub were used in immunofluorescence microscopy to demonstrate the presence of Mub on the cell surface of strain 1063. By using the antibodies in a Western blot analysis, Mub could also be detected in the growth medium. The results implicate Mub as a cell-surface protein that is involved in interactions with mucin and in colonization of the digestive tract.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): adhesion , CnBP, collagen-binding protein , MBP, maltose-binding protein , mucin and NANB, non-A, non-B
© Microbiology Society
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-148-2-433
2002-02-01
2024-04-24
Loading full text...

Full text loading...

/deliver/fulltext/micro/148/2/1480433a.html?itemId=/content/journal/micro/10.1099/00221287-148-2-433&mimeType=html&fmt=ahah

References

  1. Adlerberth I., Johansson M.-L., Molin G., Wold A. E., Ahrné S., Hanson L. Å. 1996; A mannose-specific adherence mechanism in Lactobacillus plantarum conferring binding to the human colonic cell line HT-29. Appl Environ Microbiol 62:2244–2251
     [Google Scholar]
  2. Alak J. I. B., Wolf B. W., Mdurvwa E. G., Pimentel-Smith G. E., Adeyemo O. 1997; Effect of Lactobacillus reuteri on intestinal resistance to Cryptosporidium parvum infection in a murine model of acquired immunodeficiency syndrome. J Infect Dis 175:218–221 [CrossRef]
     [Google Scholar]
  3. Aleljung P. 1994 Collagen binding of intestinal Lactobacillus reuteri: characterisation, purification and cloning PhD thesis Department of Medical Microbiology, University of Lund; Lund, Sweden:
     [Google Scholar]
  4. Aleljung P., Shen W., Rozalska B., Hellman U., Ljung Å., Wadström T. 1994; Purification of collagen binding proteins of Lactobacillus reuteri NCIB 11951 . Curr Microbiol 28:231–236 [CrossRef]
     [Google Scholar]
  5. Axelsson L., Lindgren S. 1987; Characterisation and DNA homology of Lactobacillus strains isolated from pig intestine. J Appl Bacteriol 62:433–440 [CrossRef]
     [Google Scholar]
  6. Axelsson L. T., Chung T. C., Dobrogosz W. J., Lindgren S. E. 1989; Production of a broad spectrum antimicrobial substance by Lactobacillus reuteri . Microbial Ecol Health Dis 2:131–136 [CrossRef]
     [Google Scholar]
  7. Barenkamp S. J., St Geme J. W.III. 1996; Identification of a second family of high-molecular-weight adhesion proteins expressed by non-typable Haemophilus influenzae . Mol Microbiol 19:1215–1223 [CrossRef]
     [Google Scholar]
  8. Berg R. D. 1996; The indigenous gastrointestinal microflora. Trends Microbiol 4:430–435 [CrossRef]
     [Google Scholar]
  9. Bos E. S., Schuurs A. H., van der Doelen A. A., van Roy N. 1981; 3,3′,5,5′-Tetramethylbenzidine as an Ames test negative chromogen for horse-radish peroxidase in enzyme-immunoassay. J Immunoassay 2:187–204 [CrossRef]
     [Google Scholar]
  10. Brassart D., Schiffrin E. J. 1997; The use of probiotics to reinforce mucosal defence mechanisms. Trends Food Sci Technol 8:321–326 [CrossRef]
     [Google Scholar]
  11. Braun G., McKechnie N. M., Gürr W. 1995; Molecular and immunological characterization of hr44, a human ocular component immunologically cross-reactive with antigen Ov39 of Onchocerca volvulus . J Exp Med 182:1121–1132 [CrossRef]
     [Google Scholar]
  12. Coconnier M.-H., Klaenhammer T. R., Bernet M.-F., Servin A. L., Kernèis S. 1992; Protein-mediated adhesion of Lactobacillus acidophilus BG2FO4 on human enterocyte and mucus-secreting cell line in culture. Appl Environ Microbiol 58:2034–2039
     [Google Scholar]
  13. Conway P. L., Kjelleberg S. 1989; Protein-mediated adhesion of Lactobacillus fermentum strain 737 to mouse stomach squamous epithelium. J Gen Microbiol 135:1175–1186
     [Google Scholar]
  14. Craven S. E., Williams D. D. 1998; In vitro attachment of Salmonella typhimurium to chicken cecal mucus: Effect of cations and pretreatment with Lactobacillus spp. isolated from the intestinal tract of chickens. J Food Protein 61:265–271
     [Google Scholar]
  15. Drasar B. S., Barrow P. A. 1985; The bacterial flora of the normal intestine. In Intestinal Microbiology pp 19–41 Edited by Cole J. A., Knowles C. J., Schlessinger D. Washington, DC: American Society for Microbiology;
     [Google Scholar]
  16. Edens F. W., Parkhurst C. R., Casas I. A., Dobrogosz W. J. 1997; Principles of ex ovo competitive exclusion and in ovo administration of Lactobacillus reuteri . Poultry Sci 76:179–196 [CrossRef]
     [Google Scholar]
  17. Farabaugh P. J. 1996; Programmed translational frameshifting. Microbiol Rev 60:103–134
     [Google Scholar]
  18. Felsenstein J. 1993 phylip (Phylogeny Inference Package) version 3.5c Seattle: Department of Genetics, University of Washington;
     [Google Scholar]
  19. Finegold S. M., Sutter V. L., Mathisen G. E. 1983; Normal indigenous intestinal flora. In Human Intestinal Microflora in Health and Disease pp 3–31 Edited by Hentges D. J. New York: Academic Press;
     [Google Scholar]
  20. Fischetti V. A. 2000; Surface proteins of Gram-positive bacteria. In Gram Positive Pathogens pp 11–24 Edited by Fischetti V. A., Novick R. P., Ferretti J. J., Portnoy D. A., Rood J. I. Washington, DC: American Society for Microbiology;
     [Google Scholar]
  21. Henriksson A., Conway P. L. 1992; Adhesion to porcine squamous epithelium of saccharide and protein moieties of Lactobacillus fermentum strain 104-S. J Gen Microbiol 138:2657–2661 [CrossRef]
     [Google Scholar]
  22. Jonsson H., Roos S., Ström E. 2001; Addition of mucin to the growth medium triggers mucus-binding activity in different strains of Lactobacillus reuteri in vitro . FEMS Microbiol Lett 204:19–22 [CrossRef]
     [Google Scholar]
  23. Kirjavainen P. V., Ouwehand A. C., Isolauri E., Salminen S. J. 1998; The ability of probiotic bacteria to bind to human intestinal mucus. FEMS Microbiol Lett 167:185–189 [CrossRef]
     [Google Scholar]
  24. Kotarski S. F., Savage D. C. 1979; Models for study of the specificity by which indigenous lactobacilli adhere to murine gastric epithelia. Infect Immun 63:1698–1702
     [Google Scholar]
  25. Lindgren S. E., Swaisgood H. E., Janolino V. G., Axelsson L. T., Richter C. S., Mackenzie J. M., Dobrogosz W. J. 1992; Binding of Lactobacillus reuteri to fibronectin immobilised on glass beads. Zentbl Bakteriol 277:519–528 [CrossRef]
     [Google Scholar]
  26. Molin G., Jeppsson B., Johansson M.-L., Ahrné S., Nobaek S., Ståhl M., Bengmark S. 1993; Numerical taxonomy of Lactobacillus spp. associated with healthy and diseased mucosa of the human intestines. J Appl Bacteriol 74:314–323 [CrossRef]
     [Google Scholar]
  27. Mukai T., Arihara K. 1994; Presence of intestinal lectin-binding glycoproteins on the cell surface of Lactobacillus acidophilus . Biosci Biotechnol Biochem 58:1851–1854 [CrossRef]
     [Google Scholar]
  28. Mukai T., Kaneko S., Ohori H. 1998; Haemagglutinin and glycolipid-binding activities of Lactobacillus reuteri . Lett Appl Microbiol 27:130–134 [CrossRef]
     [Google Scholar]
  29. Naito S., Hayashidani H., Kaneko K., Ogawa M., Benno Y. 1995; Development of intestinal lactobacilli in normal piglets. J Appl Bacteriol 79:230–236 [CrossRef]
     [Google Scholar]
  30. Reyes G. R., Bradley D. W., Rabin L., Fry K. E. 1990; Post-transfusion, non-A, non-B hepatitis virus and antigens. International Patent Application WO 90/00597
     [Google Scholar]
  31. Rocha C. L., Fischetti V. A. 1999; Identification and characterization of a novel fibronectin-binding protein on the surface of group A streptococci. Infect Immun 67:2720–2728
     [Google Scholar]
  32. Rojas M., Conway P. L. 1996; Colonization by lactobacilli of piglet small intestinal mucus. J Appl Bacteriol 81:474–480
     [Google Scholar]
  33. Roos S., Aleljung P., Robert N., Lee B., Lindberg M., Jonsson H., Wadström T. 1996; A collagen binding protein from Lactobacillus reuteri is part of an ABC transporter system?. FEMS Microbiol Lett 144:33–38 [CrossRef]
     [Google Scholar]
  34. Roos S., Lindgren S., Jonsson H. 1999; Autoaggregation of Lactobacillus reuteri is mediated by a putative DEAD-box helicase. Mol Microbiol 32:427–436 [CrossRef]
     [Google Scholar]
  35. Roos S., Karner F., Axelsson L., Jonsson H. 2000; Lactobacillus mucosae sp. nov., a new species with in vitro mucus-binding activity isolated from pig intestine. Int J Syst Evol Microbiol 50:251–258 [CrossRef]
     [Google Scholar]
  36. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  37. Savage D. C. 1980; Adherence of normal flora to mucosal surfaces. In Bacterial Adherence pp 33–59 Edited by Beachey E. H. London: Chapman and Hall;
     [Google Scholar]
  38. Schneewind O., Fowler A., Faull K. F. 1995; Structure of the cell wall anchor of surface proteins in Staphylococcus aureus . Science 268:103–106 [CrossRef]
     [Google Scholar]
  39. Schwartz R., Curran J. F. 1997; Analyses of frameshifting at UUU-pyrimidine sites. Nucleic Acids Res 25:2005–2011 [CrossRef]
     [Google Scholar]
  40. Shornikova A.-V., Casas I. A., Salo E., Vesikari T., Mykkänen H. 1997; Bacteriotherapy with Lactobacillus reuteri in rotavirus gastroenteritis. Pediatr Infect Dis J 16:1103–1107 [CrossRef]
     [Google Scholar]
  41. Thompson J. D., Higgins D. G., Gibson T. J. 1994; clustal w: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680 [CrossRef]
     [Google Scholar]
  42. Tuomola E. M., Ouwehand A. C., Salminen S. J. 1999; The effect of probiotic bacteria on the adhesion of pathogens to human intestinal mucus. FEMS Immunol Med Microbiol 26:137–142 [CrossRef]
     [Google Scholar]
  43. Wadström T., Anderson K., Sydow M., Axelsson L., Lindgren S., Gullmar B. 1987; Surface properties of lactobacilli isolated from the small intestine of pigs. J Appl Bacteriol 62:513–520 [CrossRef]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-148-2-433
Loading
A high-molecular-mass cell-surface protein from Lactobacillus reuteri 1063 adheres to mucus componentsThe GenBank accession number for the sequence reported in this paper is AF120104.
Microbiology 148, 433 (2002); https://doi.org/10.1099/00221287-148-2-433
/content/journal/micro/10.1099/00221287-148-2-433
/content/journal/micro/10.1099/00221287-148-2-433
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