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

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Folyl and Antifolyl Polyglutamates

Part of the book series: Advances in Experimental Medicine and Biology ((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.

This work was supported by Grant A1747 from the Natural Sciences and Engineering Research Council of Canada to E.A.C.

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Authors and Affiliations

  1. Department of Botany, University of Alberta, Edmonton, Alberta, T6G 2E9, Canada

    Edwin A. Cossins & Patrick Y. Chan

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  1. Edwin A. Cossins
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  2. Patrick Y. Chan
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Editors and Affiliations

  1. Medical College of Virginia, Virginia Commonwealth University, Richmond, Virginia, USA

    I. David Goldman

  2. National Cancer Institute, Bethesda, Maryland, USA

    Bruce A. Chabner

  3. Yale University School of Medicine, New Haven, Connecticut, USA

    Joseph R. Bertino

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© 1983 Springer Science+Business Media New York

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Cossins, E.A., Chan, P.Y. (1983). Folylpolyglutamate Synthetase Activities of Neurospora Crassa: Nature of Products Formed by Soluble and Particulate Enzymes in the Wild Type and Polyglutamate-Deficient Mutants. In: Goldman, I.D., Chabner, B.A., Bertino, J.R. (eds) Folyl and Antifolyl Polyglutamates. Advances in Experimental Medicine and Biology, vol 163. Springer, Boston, MA. https://doi.org/10.1007/978-1-4757-5241-0_15

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  • DOI: https://doi.org/10.1007/978-1-4757-5241-0_15

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4757-5243-4

  • Online ISBN: 978-1-4757-5241-0

  • eBook Packages: Springer Book Archive

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