Skip to main content
SpringerLink
Log in

Stimulation of spreading of trypsinized human fibroblasts by lysozymes fromStaphylococcus aureus, hen egg white, and human urine

  • Published:
In Vitro Aims and scope Submit manuscript

Summary

The effect of lysozyme from three different sources—Staphylococcus aureus, hen egg white, and human urine—on adhesion to substrate and spreading of trypsinized human fibroblasts was studied. Several fibroblast strains were tested under various conditions. It was found that the different cell strains did not show the same capability of spreading and stably attaching to substrates when resuspended in media not containing serum. Some strains did not spread, whereas others spread even in the absence of serum. Cell spreading in these strains did not occur when the cells were pregrown for 5 weeks in media supplemented with 1% fetal bovine serum. Lysozyme fromS. aureus allowed stable adhesion to substrate and spreading of all the fibroblast strains unable to elongate in nonsupplemented minimal essential medium. This enzyme accelerated and augmented spreading of the strains capable of elongating in the absence of serum.S. aureus lysozyme also allowed spreading and stable adhesion to substrates of all these strains when they were pregrown for 5 weeks in the presence of 1% fetal bovine serum. Furthermore, hen egg white lysozyme and the lysozyme purified from human urine were both capable of stimulating anchorage to substrate and spreading of trypsinized fibroblasts although their effect was less pronounced than that of theS. aureus lysozyme. Some tentative hypotheses for the mechanism of cell spreading in the presence of lysozyme are made. The possibility that lysozymes, virtually ubiquitous enzymes, may play a specific role in nature in the regulation of cell differentiation and tissue development is finally raised and discussed in light of several previous observations and findings.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Grinnell, F. Cellular adhesiveness and extracellular substrata. Int. Rev. Cytol. 53: 65–144; 1978.

    PubMed  CAS  Google Scholar 

  2. Wilson, A. C.; Prager, E. M. Antigenic comparison of animal lysozymes. Osserman, E. F.; Canfield, R. E.; Beychok, S. eds. Lysozyme. New York: Academic Press; 1974: 127–141.

    Google Scholar 

  3. Imoto, T.; Johnson, L. N.; North, A. C. T.; Phillips, D. C.; Rupley, J. A. Vertebrate lysozymes. Boyer, P. D. ed. The enzymes. vol. 7. New York: Academic Press; 1972: 665–868.

    Google Scholar 

  4. Jollès, P.; Bernier, I.; Berthou, J.; Charlemagne, D.; Faure, A.; Hermann, J.; Jollès, J.; Périn, J. P.; Saint-Blancard, J. From lysozymes to chitinases: structural, kinetic, and crystallographic studies. Osserman, E. F.; Canfield, R. E.; Beychok, S. eds. Lysozyme. New York: Academic Press; 1974: 31–54.

    Google Scholar 

  5. Chipman, D. M.; Sharon, N. Mechanism of lysozyme action. Science 165: 454–465; 1969.

    Article  PubMed  CAS  Google Scholar 

  6. Smith, H. Survival of vegetative bacteria in animals. Symp. Soc. Gen. Microbiol. 26: 299–326; 1976.

    Google Scholar 

  7. Osserman, E. F. Preface. Osserman, E. F.; Canfield, R. E.; Beychok, S. eds. Lysozyme. New York: Academic Press; 1974: xxv-xxvi.

    Google Scholar 

  8. Osserman, E. F.; Klockars, M.; Halper, J.; Fischel, R. E. Effects of lysozyme on normal and transformed cells. Nature 243: 331–335; 1973.

    Article  PubMed  CAS  Google Scholar 

  9. Asdourian, H.; Chu, L.; Lau, K.; Amos, H. Lysozyme: evidence for effects on chick fibroblasts, HeLa cells, and their products. Biochem. Biophys. Res. Commun. 64: 1144–1151; 1975.

    Article  Google Scholar 

  10. Satta, G.; Varaldo, P. E.; Azzarone, B. Stimulation of DNA synthesis and cell proliferation of transformed human fibroblasts by pureStaphylococcus aureus lysozyme. Microbios Lett. 6: 55–60; 1978.

    Google Scholar 

  11. Satta, G.; Varaldo, P. E.; Azzarone, B.; Romanzi, C. A. Effects ofStaphylococcus aureus lysozyme on human fibroblasts. Cell Biol. Int. Rep. 3: 525–533; 1979.

    Article  PubMed  CAS  Google Scholar 

  12. Atherly, A. G.; Barnhart, B. J.; Kramer, P. M. Growth and biochemical characteristics of a detachment variant of CHO cells. J. Cell. Physiol. 90: 375–386; 1977.

    Article  PubMed  CAS  Google Scholar 

  13. Pouysségur, J.; Willingham, M.; Pastan, I. Role of cell surface carbohydrates and proteins in cell behavior: studies on the biochemical reversion of anN-acetylglucosamine-deficient fibroblast mutant. Proc. Natl. Acad. Sci. U.S.A. 74: 243–247; 1977.

    Article  PubMed  Google Scholar 

  14. Hynes, R. O. Cell surface protein and malignant transformation. Biochim. Biophys. Acta 458: 73–107; 1976.

    PubMed  CAS  Google Scholar 

  15. Roseman, S. Sugars of the cell membrane. Hosp. Pract. 10, no. 1: 61–71; 1975.

    Google Scholar 

  16. Osserman, E. F.; Lawlor, D. P. Serum and urinary lysozyme (muramidase) in monocytic and monomyelocytic leukemia. J. Exp. Med. 124: 921–952; 1966.

    Article  PubMed  CAS  Google Scholar 

  17. Canfield, R. E.; Collins, J. C.; Sobel, J. H. Human leukemia lysozyme. Osserman, E. F.; Canfield, R. E.; Beychok, S. eds. Lysozyme. New York: Academic Press; 1974: 63–70.

    Google Scholar 

  18. Rupley, J. A. The hydrolysis of chitin by concentrated hydrochloric acid, and the preparation of low-molecular-weight substrates for lysozyme. Biochim. Biophys. Acta 83: 245–255; 1964.

    PubMed  CAS  Google Scholar 

  19. Fahey, J. L.; Terry, E. W. Ion exchange chromatography and gel filtration. Weir, D. M. ed. Handbook of experimental immunology. vol. 1. Oxford: Blackwell; 1978: 8.1–8.16.

    Google Scholar 

  20. Taylor, A. C. Attachment and spreading of cells in culture. Exp. Cell Res., Suppl. 8: 154–173; 1961.

    Article  PubMed  Google Scholar 

  21. Wolpert, L.; Macpherson, I.; Todd, I. Cell spreading and cell movement: an active or passive process? Nature 223: 513–514; 1969.

    Article  Google Scholar 

  22. Grinnell, F. Cell spreading factor. Occurrence and specificity of action. Exp. Cell Res. 102: 51–62; 1976.

    Article  PubMed  CAS  Google Scholar 

  23. Culp, L. A.; Buniel, J. F. Substrate-attached serum and cell proteins in adhesion of mouse fibroblasts. J. Cell. Physiol. 88: 89–106; 1976.

    Article  PubMed  CAS  Google Scholar 

  24. Witkowski, J. A.; Brighton, W. D. Stages of spreading of human diploid cells on glass surfaces. Exp. Cell Res. 68: 372–380; 1971.

    Article  PubMed  CAS  Google Scholar 

  25. Rajaraman, R.; Rounds, D. E.; Yen, S. P. S.; Rembaum, A. A scanning electron microscope study of cell adhesion and spreading in vitro. Exp. Cell Res. 88: 327–339; 1974.

    Article  PubMed  CAS  Google Scholar 

  26. Grinnell, F.; Minter, D. Attachment and spreading of baby hamster kidney cells to collagen substrate: effects of cold-insoluble globulin. Proc. Natl. Acad. Sci. U.S.A. 75: 4408–4412; 1978.

    Article  PubMed  CAS  Google Scholar 

  27. Irie, R. F.; Irie, K.; Morton, D. L. Natural antibody in human serum to a neoantigen in human cultured cells grown in fetal bovine serum. J. Natl. Cancer Inst. 52: 1051–1058; 1974.

    PubMed  CAS  Google Scholar 

  28. Wadström, T.; Hisatsune, K. Bacteriolytic enzymes fromStaphylococcus aureus. Purification of an endo-β-N-acetylglucosaminidase. Biochem. J. 120: 725–734; 1970.

    PubMed  Google Scholar 

  29. Wadström, T. Bacteriolytic enzymes fromStaphylococcus aureus. Properties of the endo-β-N-acetylglucosaminidase. Biochem. J. 120: 745–752; 1970.

    PubMed  Google Scholar 

  30. Wadström, T.; Vesterberg, O. Studies on endo-β-N-acetylglucosaminidase, staphylolytic peptidase, andN-acetylmuramyl-l-alanine amidase in lysostaphin and fromStaphylococcus aureus. Acta Pathol. Microbiol. Scand. Sect. B 79: 248–264; 1971.

    Google Scholar 

  31. Iversen, O. J.; Grov, A. Studies on lysostaphin. Separation and characterization of three enzymes. Eur. J. Biochem. 38: 293–300; 1973.

    Article  PubMed  CAS  Google Scholar 

  32. Ui, N. Isoelectric points and conformation of proteins. I. Effect of urea on the behaviour of some proteins in isoelectric focusing. Biochim. Biophys. Acta 229: 567–581; 1971.

    PubMed  CAS  Google Scholar 

  33. Vesterberg, O.; Wadström, T.; Vesterberg, K.; Svensson, H.; Malmgren, B. Studies of extracellular proteins fromStaphylococcus aureus. I. Separation and characterization of enzymes and toxins by isoelectric focusing. Biochim. Biophys. Acta 133: 435–445; 1967.

    PubMed  CAS  Google Scholar 

  34. Aburthnott, J. P. Staphylococcal toxins. Schlessinger, D., ed. Microbiology-1975. Washington: American Society for Microbiology; 1975: 267–271.

    Google Scholar 

  35. Wadström, T.; Hisatsune, K. Bacteriolytic enzymes fromStaphylococcus aureus. Specificity of action of endo-β-N-acetylglucosaminidase. Biochem. J. 120: 735–744; 1970.

    PubMed  Google Scholar 

  36. Kuettner, K. E.; Soble, L. W.; Eisenstein, R.; Yaeger, J. A. The influence of lysozyme on the appearance of epiphyseal cartilage in organ culture. Calcif. Tissue Res. 2: 93–105; 1968.

    Article  PubMed  CAS  Google Scholar 

  37. Kuettner, K. E.; Soble, L. W.; Guenther, H. L.; Croxen, R. L.; Eisenstein, R. Lysozyme in epiphyseal cartilage. I. The nature of the morphologic response of cartilage in culture to exogenous lysozyme. Calcif. Tissue Res. 5: 56–63; 1970.

    Article  PubMed  CAS  Google Scholar 

  38. Takaoka, T.; Katsuta, H.; Ishiki, S.; Konishi, T. Effects of lysozyme on the proliferation of fibroblasts in tissue culture. Jpn. J. Exp. Med. 42: 221–232; 1972.

    PubMed  CAS  Google Scholar 

  39. Osserman, E. F.; Klockars, M.; Halper, J.; Fischel, R. E. Studies of the effects of lysozyme on mammalian cells. Osserman, E. F.; Canfield, R. E.; Beychok, S. eds. Lysozyme. New York: Academic Press; 1974: 471–490.

    Google Scholar 

  40. Adinolfi, M.; Loeb, J. N.; Osserman, E. F. Agglutination of rat liver mitochondria by lysozyme. Osserman, E. F.; Canfield, R. E.; Beychok, S. eds. Lysozyme. New York: Academic Press; 1974: 463–470.

    Google Scholar 

  41. Daneo-Moore, L.; Shockman, G. D. The bacterial cell surface in growth and division. Poste, G.; Nicolson, G. L. eds. The synthesis, assembly and turnover of cell surface components. Cell surface reviews. vol. 4. Amsterdam: North-Holland, 1977: 597–715.

    Google Scholar 

  42. Rogers, H. J. Bacterial growth and the cell envelope. Bacteriol. Rev. 61: 155–171; 1970.

    CAS  Google Scholar 

  43. Thompson, J. S. Role of autolytic enzymes in the growth and morphogenesis of bacterial cell walls. J. Theor. Biol. 33: 63–75; 1971.

    Article  PubMed  CAS  Google Scholar 

  44. Klockars, M.; Reitamo, S.; Adinolfi, M. Ontogeny of human lysozyme. Distribution in fetal tissues. Biol. Neonat 32: 243–249; 1977.

    CAS  Google Scholar 

  45. Kuettner, K. E.; Guenther, H. L.; Ray, R. D.; Schumacher, G. F. B. Lysozyme in preosseous cartilage. Calcif. Tissue Res. 1: 298–305; 1968.

    Article  PubMed  CAS  Google Scholar 

  46. Kuettner, K. E.; Sorgente, N.; Croxen, R. L.; Howell, D. S.; Pita, J. C. Lysozyme in preosseous cartilage. VII. Evidence for physiological role of lysozyme in normal and endochondral calcification. Biochim. Biophys. Acta 373: 335–344; 1974.

    Google Scholar 

  47. Roberts, R. M.; Baxer, F. W.; Baldwin, N.; Pollard, W. E. Progesterone induction of lysozyme and peptidase activities in the porcine uterus. Arch. Biochem. Biophys. 177: 499–507; 1976.

    Article  PubMed  CAS  Google Scholar 

  48. Klockars, M.; Reitamo, S. Tissue distribution of lysozyme in man. J. Histochem. Cytochem. 23: 932–940; 1975.

    PubMed  CAS  Google Scholar 

  49. Varaldo, P. E.; Satta, G.; Mingari, M. C.; Romanzi, C. A. Effects ofStaphylococcus aureus lysozyme on the immune competent cells. Proc. XII Int. Congr. Microbiol., Munich, September 3–8, 1978.

  50. Avery, R. J.; Levy, J. A. Relationship of endogenous murine xenotropic type C virus production to spontaneous transformation of cultured cells. J. Gen. Virol. 39: 427–435; 1978.

    Article  PubMed  CAS  Google Scholar 

  51. Levy, J. A.; Kazan, P.; Varnier, O.; Kleiman, H. Murine xenotropic type C viruses. I. Distribution and further characterization of the virus in NZB mice. J. Virol. 16: 844–853; 1975.

    PubMed  CAS  Google Scholar 

  52. Yamada, K. M.; Olden, K. Fibronectins—adhesive glycoproteins of cell surface and blood. Nature 275: 179–184; 1978.

    Article  PubMed  CAS  Google Scholar 

  53. Grinnell, F. The serum dependence of baby hamster kidney cell attachment to a substratum. Exp. Cell Res. 97: 265–274; 1976.

    Article  PubMed  CAS  Google Scholar 

  54. Rees, D. A.; Lloyd, W. C.; Thom, D. Control of grip and stick in cell adhesion through lateral relationships of membrane glycoproteins. Nature 267: 124–128; 1977.

    Article  PubMed  CAS  Google Scholar 

  55. Levi, A.; Cimino, M.; Mercanti, D.; Chen, J. S.; Calissano, P. Interaction of nerve growth factor with tubulin. Studies on binding and induced polymerization. Biochim. Biophys. Acta 399: 50–60; 1975.

    PubMed  CAS  Google Scholar 

  56. Nicolson, G. L. Transmembrane control of the receptors on normal and tumor cells. I. Cytoplasmic influence over cell surface components. Biochim. Biophys. Acta 457: 57–108; 1976.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Istituto di Microbiologia dell’Università di Genova, Viale Benedetto XV, 10, 16132, Genova, Italy

    G. Satta, B. Azzarone, P. E. Varaldo, R. Fontana & S. Valisena

Authors
  1. G. Satta
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. B. Azzarone
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. P. E. Varaldo
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. R. Fontana
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. S. Valisena
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

This work was supported by Grant 79.00677.96 of Piano Finalizzato Tumori by the Consiglio Nazionale delle Ricerche of Italy.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Satta, G., Azzarone, B., Varaldo, P.E. et al. Stimulation of spreading of trypsinized human fibroblasts by lysozymes fromStaphylococcus aureus, hen egg white, and human urine. In Vitro 16, 738–750 (1980). https://doi.org/10.1007/BF02619307

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02619307

Key words

Search

Navigation