Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1981 Mar;78(3):1495–1498. doi: 10.1073/pnas.78.3.1495

15N NMR studies of nitrogen metabolism in intact mycelia of Neurospora crassa.

T L Legerton, K Kanamori, R L Weiss, J D Roberts
PMCID: PMC319157  PMID: 6453347

Abstract

Suspensions of intact mycelia of Neurospora crassa grown in medium containing 15NH4Cl have been found to give well-resolved 15N nuclear magnetic resonance spectra for metabolites that play crucial roles in intermediary nitrogen metabolism. These include the amide nitrogen of glutamine, the alpha-amino nitrogens of glutamate and other amino acids, the guanidino nitrogens of arginine, the ureido nitrogen of citrulline, the side-chain nitrogens of ornithine or lysine, or both, and uridine diphosphates. The turnover time of glutamine in vivo was estimated to be less than 1 hr by using nuclear magnetic resonance spectroscopy in conjunction with tracer methodologies. Applications of these techniques to the study of nitrogen metabolism are discussed.

Full text

PDF
1495

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Basabe J. R., Lee C. A., Weiss R. L. Enzyme assays using permeabilized cells of Neurospora. Anal Biochem. 1979 Jan 15;92(2):356–360. doi: 10.1016/0003-2697(79)90670-5. [DOI] [PubMed] [Google Scholar]
  2. Botto R. E., Roberts J. D. Nitrogen-15 magnetic resonance spectroscopy. Natural-abundance nitrogen-15 spectra of some 2-amino-2-deoxy-D-hexose derivatives. J Org Chem. 1977 Jun 24;42(13):2247–2249. doi: 10.1021/jo00433a012. [DOI] [PubMed] [Google Scholar]
  3. DE TERRA N., TATUM E. L. Colonial growth of Neurospora. Sorbose and enzymes alter the composition of the cell wall and induce morphological changes. Science. 1961 Oct 13;134(3485):1066–1068. doi: 10.1126/science.134.3485.1066. [DOI] [PubMed] [Google Scholar]
  4. Gust D., Moon R. B., Roberts J. D. Applications of natural-abundance nitrogen-15 nuclear magnetic resonance to large biochemically important molecules. Proc Natl Acad Sci U S A. 1975 Dec;72(12):4696–4700. doi: 10.1073/pnas.72.12.4696. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hubbard J. S., Stadtman E. R. Regulation of glutamine synthetase. II. Patterns of feedback inhibition in microorganisms. J Bacteriol. 1967 Mar;93(3):1045–1055. doi: 10.1128/jb.93.3.1045-1055.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Karlin J. N., Bowman B. J., Davis R. H. Compartmental behavior of ornithine in Neurospora crassa. J Biol Chem. 1976 Jul 10;251(13):3948–3955. [PubMed] [Google Scholar]
  7. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  8. Lapidot A., Irving C. S. Comparative in vivo nitrogen-15 nuclear magnetic resonance study of the cell wall components of five Gram-positive bacteria. Biochemistry. 1979 Feb 20;18(4):704–714. doi: 10.1021/bi00571a024. [DOI] [PubMed] [Google Scholar]
  9. Lapidot A., Irving C. S. Dynamic structure of whole cells probed by nuclear Overhauser enhanced nitrogen-15 nuclear magnetic resonance spectroscopy. Proc Natl Acad Sci U S A. 1977 May;74(5):1988–1992. doi: 10.1073/pnas.74.5.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lapidot A., Irving C. S. Nitrogen-15 and carbon-13 dynamic nuclear magnetic resonance study of chain segmental motion of the peptidoglycan pentaglycine chain of 15N-Gly- and 13C2-Gly-labeled Staphylococcus aureus cells and isolated cell walls. Biochemistry. 1979 May 1;18(9):1788–1796. doi: 10.1021/bi00576a024. [DOI] [PubMed] [Google Scholar]
  11. Llinás M., Wüthrich K., Schwotzer W., Von Philipsborn W. 15N nuclear magnetic resonance of living cells. Nature. 1975 Oct 30;257(5529):817–818. doi: 10.1038/257817a0. [DOI] [PubMed] [Google Scholar]
  12. Markowski V., Sullivan G. R., Roberts J. D. Nitrogen-15 nuclear magnetic resonance spectroscopy of some nucleosides and nucleotides. J Am Chem Soc. 1977 Feb 2;99(3):714–718. doi: 10.1021/ja00445a009. [DOI] [PubMed] [Google Scholar]
  13. Martegani E., Alberghina L. Intracellular protein degradation in Neurospora crassa. J Biol Chem. 1979 Aug 10;254(15):7047–7054. [PubMed] [Google Scholar]
  14. Moon R. B., Richards J. H. Determination of intracellular pH by 31P magnetic resonance. J Biol Chem. 1973 Oct 25;248(20):7276–7278. [PubMed] [Google Scholar]
  15. Navon G., Shulman R. G., Yamane T., Eccleshall T. R., Lam K. B., Baronofsky J. J., Marmur J. Phosphorus-31 nuclear magnetic resonance studies of wild-type and glycolytic pathway mutants of Saccharomyces cerevisiae. Biochemistry. 1979 Oct 16;18(21):4487–4499. doi: 10.1021/bi00588a006. [DOI] [PubMed] [Google Scholar]
  16. SMITH E. J., WHEAT R. W. Acid-soluble nucleotide spectrum of Neurospora crassa. Arch Biochem Biophys. 1960 Feb;86:267–269. doi: 10.1016/0003-9861(60)90416-1. [DOI] [PubMed] [Google Scholar]
  17. Schaefer J., Stejskal E. O., McKay R. A. Cross-polarization NMR of N-15 labeled soybeans. Biochem Biophys Res Commun. 1979 May 14;88(1):274–280. doi: 10.1016/0006-291x(79)91726-1. [DOI] [PubMed] [Google Scholar]
  18. Shulman R. G., Brown T. R., Ugurbil K., Ogawa S., Cohen S. M., den Hollander J. A. Cellular applications of 31P and 13C nuclear magnetic resonance. Science. 1979 Jul 13;205(4402):160–166. doi: 10.1126/science.36664. [DOI] [PubMed] [Google Scholar]
  19. Weiss R. L., Anterasian G. P. Control of arginine metabolism in Neurospora. Induction of ornithine aminotransferase. J Biol Chem. 1977 Oct 25;252(20):6974–6980. [PubMed] [Google Scholar]
  20. Weiss R. L., Davis R. H. Intracellular localization of enzymes of arginine metabolism in Neurospora. J Biol Chem. 1973 Aug 10;248(15):5403–5408. [PubMed] [Google Scholar]
  21. Weiss R. L. Intracellular localization of ornithine and arginine pools in Neurospora. J Biol Chem. 1973 Aug 10;248(15):5409–5413. [PubMed] [Google Scholar]
  22. Wolf E. C., Weiss R. L. Acetylglutamate kinase. A mitochondrial feedback-sensitive enzyme of arginine biosynthesis in Neurospora crassa. J Biol Chem. 1980 Oct 10;255(19):9189–9195. [PubMed] [Google Scholar]
  23. den Hollander J. A., Brown T. R., Ugurbil K., Shulman R. G. 13C nuclear magnetic resonance studies of anaerobic glycolysis in suspensions of yeast cells. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6096–6100. doi: 10.1073/pnas.76.12.6096. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES