Advertisement

Folylpolyglutamate Synthetase Activities of Neurospora Crassa: Nature of Products Formed by Soluble and Particulate Enzymes in the Wild Type and Polyglutamate-Deficient Mutants

  • Edwin A. Cossins
  • Patrick Y. Chan
Chapter
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 163)

Abstract

The folylpolyglutamate synthetase activities of Neurospora crassa wild type (FGSC 853) and two polyglutamate-deficient mutants (met-6, 35809, FGSC 1330 and mac, 65108, FGSC 3609) were examined using dialyzed extracts prepared during exponential mycelial growth. Enzyme assay was based on incorporation of [U-3H]glutamate in folyl-polyglutamates that were separated by gradient elution from DEAE-cellulose. Extracts of the wild type produced H4PteGlu2 (15%), H4PteGlu3 (35%) and H4PteGlu6 (50%) when anaerobically incubated with glutamate, ATP, and H4PteGlu. Under these conditions, the met-6 produced only H4PteGlu2 and higher polyglutamates (H4PteGlu4 and H4PteGlu5) were not utilized. The mac mutant failed to catalyze addition of glutamate to H4PteGlu. However, H4PteGlu2 was effectively converted to the tri-, and hexaglutamates. Mixing wild type and met-6 protein stimulated the formation of tri-, and hexaglutamates. Mixing mac and met-6 extracts resulted in H4PteGlua and H4PteGlu6 labeling when glutamate and H4PteGlu were provided.

Keywords

Potassium Phosphate Wild Type Protein Neurospora CRASSA Synthetase Activity 2S04 Fraction 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
This is a preview of subscription content, log in to check access.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Gawthorne, J. M., and Smith, R. M., Biochem. J., 136, 295–301 (1973).PubMedGoogle Scholar
  2. 2.
    Taylor, R. T., and Hanna, M. L., Arch. Biochem. Biophys., 181, 331–344 (1977).PubMedCrossRefGoogle Scholar
  3. 3.
    Taylor, R. T., and Hanna, M. L., Arch. Biochem. Biophys., 197, 36–43 (1979).PubMedCrossRefGoogle Scholar
  4. 4.
    McGuire, J. J., Kitamoto, Y., Hsieh, P., Coward, J. K., and Bertino, J. R., in: Chemistry and Biology of Pteridines (R. L. Kisliuk and G. M. Brown, eds.), pp. 471–476, Elsevier-North Holland, New York (1979).Google Scholar
  5. 5.
    Masurekar, M., and Brown, G. M., Biochemistry, 14, 2424–2430 (1975).PubMedCrossRefGoogle Scholar
  6. 6.
    Shane, B., Brody, T., and Stokstad, E. L. R., in: Chemistry and Biology of Pteridines (R. L. Kisliuk, and G. M. Brown, eds.), pp. 341–346, Elsevier-North Holland, New York (1979).Google Scholar
  7. 7.
    Shane, B., J. Biol. Chem., 255, 5655–5662 (1980).PubMedGoogle Scholar
  8. 8.
    Shane, B., J. Biol. Chem., 255, 5663–5667 (1980).PubMedGoogle Scholar
  9. 9.
    Schetel, M. F., Boehue, J. W., and Libby, D. A., J. Biol. Chem., 240, 3154–3158 (1965).Google Scholar
  10. 10.
    Lor, K. L., and Cossins, E. A., Biochem. J., 130, 773–783 (1972).PubMedGoogle Scholar
  11. 11.
    Baugh, C. M., Braverman, E., and Nair, M. G., in: Chemistry and Biology of Pteridines (W. Pfleiderer, ed.), pp. 465–474, Walter de Gruyter, Berlin (1975).Google Scholar
  12. 12.
    Bassett, J. R., Weir, D. G., and Scott, J. M., J. Gen. Microbiol., 93, 169–172 (1976).PubMedCrossRefGoogle Scholar
  13. 13.
    Cossins, E. A., Chan, P.-Y., and Combepine, G., Biochem. J., 160, 305–314 (1976).PubMedGoogle Scholar
  14. 14.
    Cossins, E. A., and Chan, P. Y., Plant Biochemical J. S. M. Sircar Mem. Vol., 53–62 (1980).Google Scholar
  15. 15.
    Chan, P. Y., and Cossins, E. A., Arch. Biochem. Biophys., 200, 346–356 (1980).PubMedCrossRefGoogle Scholar
  16. 16.
    Sakami, W., Ritari, S. J., Black, C. W., and Rzepka, J., Fed. Proc, 32, 471 (1973).Google Scholar
  17. 17.
    Ritari, S. J., Sakami, W., Black, C. W., and Rzepka, J. Anal. Biochem., 63, 118–129 (1975).PubMedCrossRefGoogle Scholar
  18. 18.
    Ritari, S. J., Sakami, W., Black, C. W., and Rzepka, J., Neuro-spora Newletter, 20, 26–27 (1973).Google Scholar
  19. 19.
    Ritari, S. J., Sakami, W., and Black, C. W., Neurospora New-letter, 20, 27 (1973).Google Scholar
  20. 20.
    Reid, V. E., and Friedkin, M., Mol. Pharmacol., 9, 74–80 (1973).PubMedGoogle Scholar
  21. 21.
    Vogel, J. H., Amer. Natur., 98, 435–446 (1964).CrossRefGoogle Scholar
  22. 22.
    Kuntzel, H., and Noll, H., Nature, 215, 1340–1345 (1967).PubMedCrossRefGoogle Scholar
  23. 23.
    Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J., J. Biol. Chem., 193, 265–275 (1951).PubMedGoogle Scholar
  24. 24.
    Clandinin, M. T., and Cossins, E. A., Biochem. J., 128, 29–40 (1972).PubMedGoogle Scholar
  25. 25.
    Turner, J. C, Int. J. Appl. Radiat. Isotop., 20, 499–505 (1969).CrossRefGoogle Scholar
  26. 26.
    Covey, J. M., Life Sciences, 26, 665–678 (1980).PubMedCrossRefGoogle Scholar
  27. 27.
    Cossins, E. A., in: The Biochemistry of Plants, Vol. 2, (D. D. Davis, ed.), pp. 365–418, Academic Press, New York (1980).Google Scholar
  28. 28.
    Zelikson, R., and Luzzati, M., Eur. J. Biochem., 79, 285–292 (1977).PubMedCrossRefGoogle Scholar
  29. 29.
    Murray, N. E., Genetics, 61, 67–77 (1969).PubMedGoogle Scholar
  30. 30.
    Burton, E., Selhub, J., and Sakami, W., Biochem. J., 111, 793–795 (1969).PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1983

Authors and Affiliations

  • Edwin A. Cossins
    • 1
  • Patrick Y. Chan
    • 1
  1. 1.Department of BotanyUniversity of AlbertaEdmontonCanada

Personalised recommendations

Cite chapter

Buy options