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.
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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

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