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Archives of Microbiology

, Volume 118, Issue 1, pp 55–60 | Cite as

Regulation of proteolytic enzymes in axenically grown myxamoebae of Physarum polycephalum

  • Annegret Haars
  • Clare H. R. McCullough
  • Aloys Hüttermann
  • Jennifer Dee
Article

Abstract

The cultivation of Physarum polycephalum amoebae in two media with different protein contents revealed a regulation of aminopeptidases and proteases depending on the albumin content of the medium: in growing amoebae and plasmodia the aminopeptidases have similar isoenzyme patterns and relative activities against nitroanilides. One alanine and four leucine aminopeptidase isoenzymes were found within the slightly acid pH range. During growth amoebae secrete—different from plasmodia—leucine aminopeptidase into the medium with low protein content. In an albumin-rich medium additional alanine aminopeptidase activity was found. Out of nine plasmodial proteases four were found in amoebae too. Only one band (pI 3.6) was present in the protein-poor medium. No protease activity could be detected in the proteinrich medium.

Key words

Physarum polycephalum Amoebae Aminopeptidases Acid proteases Regulation Development Differential gene activity 

Abbreviations

BSA

bovine serum albumin

SD

semi-defined (medium with low protein content; Table 1)

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References

  1. Brand, G., Hüttermann, A., Haugli, F. B.: Differential expression of RNase activities in the life cycle of Physarum polycephalum. Naturwissenschaften 11, 535–536 (1975)Google Scholar
  2. Cooke, D. J., Dee, J.: Methods for the isolation and analysis of plasmodial mutants in Physarum polycephalum. Genet. Res. 24, 175–187 (1975)Google Scholar
  3. Daniel, J. W., Baldwin, H. H.: Methods of culture for plasmodial Myxomycetes. In: Methods in cell physiology, Vol. 1 (D. M. Prescott, ed.), pp. 9–41. New York: Academic Press 1964Google Scholar
  4. Dee, J.: Slime moulds in biological research. Sci. Prog. (Oxf.) 62, 523–542 (1975)Google Scholar
  5. Drucker, H.: Regulation of exocellular proteases in Neurospora crassa: metabolic requirements of the process. J. Bacteriol. 122, 1117–1125 (1975)PubMedGoogle Scholar
  6. Ellman, G. L.: The biuret reaction: changes in the ultraviolet absorption spectra and its application to the determination of peptide bonds. Analyt. Biochem. 3, 40–48 (1962)PubMedGoogle Scholar
  7. Farr, D. R., Horisberger, M., Jollès, P.: An extracellular rennin-like enzyme produced by Physarum polycephalum. Biochim. Biophys. Acta 334, 410–416 (1974)Google Scholar
  8. Fawcett, J. K., Scott, J. E.: A rapid and precise method for the determination of urea. J. Clin. Pathol. 13, 156–159 (1960)PubMedGoogle Scholar
  9. Goodman, E. M.: Axenic culture of myxamoebae of the myxomycete Physarum polycephalum. J. Bacteriol. 111, 242–247 (1972)PubMedGoogle Scholar
  10. Haars, A.: Entwicklungsgenetische Untersuchungen an Physarum polycephalum: Probleme bei der Selektion von Enzym-Mutanten bei Myxamöben. Ph.D. thesis. Univ. Göttingen (1976)Google Scholar
  11. Hanson, M. A., Marzluff, G. A.: Regulation of a sulfur-controlled protease in Neurospora crassa. J. Bacteriol. 116, 785–789 (1973)PubMedGoogle Scholar
  12. Haugli, F. B.: Mutagenesis, selection and genetic analysis in Physarum polycephalum. Ph.D. thesis, Univ. Wisconsin (1971)Google Scholar
  13. Henney, H. R., Jun., Henney, M. R.: Nutritional requirements for the growth in pure culture of the myxomycete Physarum rigidum and related species. J. Gen. Microbiol. 53, 333–339 (1968)Google Scholar
  14. Hoffmann, W.: Entwicklungsgenetische Untersuchungen an Physarum polycephalum: Aktivität, Isoenzymmuster und differentielle Synthese von Aminopeptidasen während der Sklerotienbildung. Ph.D thesis, Univ. Tübingen (1975)Google Scholar
  15. Hoffmann, W., Hüttermann, A.: Aminopeptidases of Physarum polycephalum. Activity, isoenzyme pattern, and synthesis during differentiation. J. Biol. Chem. 250, 7420–7427 (1975)PubMedGoogle Scholar
  16. Hüttermann, A.: Physarum polycephalum, Studienobjekt der Zellbiologie. Ber. Dtsch. Bot. Ges. 86, 165–179 (1973)Google Scholar
  17. Kint, J. A.: A simple microtechnique for isoelectric focusing on a density gradient. Anal. Biochem. 67, 679–683 (1975)PubMedGoogle Scholar
  18. LeStourgeon, W. M., Goodman, E. M., Rusch, H. P.: The nuclear acidic proteins from haploid and diploid cell states of Physarum polycephalum. Biochim. Biophys. Acta 317, 524–528 (1973)Google Scholar
  19. Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurement with the Folin-phenol reagent. J. Biol. Chem. 193, 265–275 (1951)PubMedGoogle Scholar
  20. McCullough, C. H. R., Dee, J.: Defined and semi-defined media for the growth of amoebae of Physarum polycephalum. J. Gen. Microbiol. 95, 151–158 (1976)PubMedGoogle Scholar
  21. Panyim, S., Chalkley, R.: High resolution acrylamide gel electrophoresis of histones. Arch. Biochem. Biophys. 130, 337–342 (1969)PubMedGoogle Scholar
  22. Peters, T., Jr., Hawn, C.: Isolation of two large peptide fragments from the amino- and carboxyl-terminal positions of bovine serum albumin. J. Biol. Chem. 242, 1566–1573 (1967)PubMedGoogle Scholar
  23. Rosen, H.: A modified ninhydrin colorimetric analysis for amino acids. Arch. Biochem. Biophys. 67, 10–15 (1957)PubMedGoogle Scholar
  24. Tomarelli, R. M., Charney, J., Harding, M. L.: The use of azoalbumin as a substrate in the colorimetric determination of peptic and tryptic activity. J. Lab. Clin. Med. 34, 428–433 (1949)Google Scholar

Copyright information

© Springer-Verlag 1978

Authors and Affiliations

  • Annegret Haars
    • 1
  • Clare H. R. McCullough
    • 2
  • Aloys Hüttermann
    • 1
  • Jennifer Dee
    • 2
  1. 1.Forstbotanisches Institut der Universität GöttingenGöttingenFederal Republic of Germany
  2. 2.Department of GeneticsThe UniversityLeicesterEngland

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