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. 1983 Aug 15;214(2):345–351. doi: 10.1042/bj2140345

Polyamine-stimulated alteration of the ornithine decarboxylase molecule in Physarum polycephalum.

J L Mitchell, J M Wilson
PMCID: PMC1152254  PMID: 6615477

Abstract

The molecular mechanism for polyamine-stimulated feedback modification of ornithine decarboxylase isolated from Physarum polycephalum was investigated by using two-dimensional polyacrylamide-gel electrophoresis. Partially purified A-form enzyme was converted into the B-form enzyme by isolated fractions of the Physarum A-B-converting protein, and the substrates and products were subsequently labelled by covalent addition of alpha-difluoro[14C]methylornithine, an enzyme-activated irreversible inhibitor. The active (A-form) and inactive (B-form) states of this enzyme were found to have the same Mr value, 52 000, yet they differed noticeably in their pI values, 5.45 and 5.65 respectively. In further experiments, the use of high-specific-radioactivity [3H]spermidine to stimulate this enzyme modification was shown not to result in the covalent attachment of this polyamine to ornithine decarboxylase. These results demonstrate that the polyamine-induced modification of ornithine decarboxylase in Physarum is not due to any of the mechanisms previously suggested for ornithine decarboxylase inactivation in this and other eukaryotes, namely phosphorylation, covalent polyamine addition or the non-covalent association of a specific low-Mr protein.

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Selected References

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

  1. Branca A. A., Herbst E. J. Inhibition of ornithine decarboxylase of HeLa cells by diamines and polyamines. Effect on cell proliferation. Biochem J. 1980 Mar 15;186(3):925–931. doi: 10.1042/bj1860925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Clark J. L., Fuller J. L. Regulation of ornithine decarboxylase in 3T3 cells by putrescine and spermidine: indirect evidence for translational control. Biochemistry. 1975 Oct 7;14(20):4403–4409. doi: 10.1021/bi00691a010. [DOI] [PubMed] [Google Scholar]
  3. Heby O. Role of polyamines in the control of cell proliferation and differentiation. Differentiation. 1981;19(1):1–20. doi: 10.1111/j.1432-0436.1981.tb01123.x. [DOI] [PubMed] [Google Scholar]
  4. Heller J. S., Canellakis E. S. Cellular control of ornithine decarboxylase activity by its antizyme. J Cell Physiol. 1981 May;107(2):209–217. doi: 10.1002/jcp.1041070206. [DOI] [PubMed] [Google Scholar]
  5. Heller J. S., Fong W. F., Canellakis E. S. Induction of a protein inhibitor to ornithine decarboxylase by the end products of its reaction. Proc Natl Acad Sci U S A. 1976 Jun;73(6):1858–1862. doi: 10.1073/pnas.73.6.1858. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kallio A. Evidence for several mechanisms involved in the regulation of ornithine decarboxylase by diamines during rat liver regeneration. Acta Chem Scand B. 1978;B32(10):759–764. doi: 10.3891/acta.chem.scand.32b-0759. [DOI] [PubMed] [Google Scholar]
  7. Kallio A., Löfman M., Pösö H., Jänne J. Inhibition of ornithine decarboxylase by diamines in regenerating rat liver. Evidence for direct action on the accumulation of the enzyme protein. FEBS Lett. 1977 Jul 1;79(1):195–199. [PubMed] [Google Scholar]
  8. McCann P. P., Tardif C., Hornsperger J. M., Böhlen P. Two distinct mechanisms for ornithine decarboxylase regulation by polyamines in rat hepatoma cells. J Cell Physiol. 1979 May;99(2):183–190. doi: 10.1002/jcp.1040990204. [DOI] [PubMed] [Google Scholar]
  9. McCann P. P., Tardif C., Mamont P. S. Regulation of ornithine decarboxylase by ODC-antizyme in HTC cells. Biochem Biophys Res Commun. 1977 Apr 25;75(4):948–954. doi: 10.1016/0006-291x(77)91474-7. [DOI] [PubMed] [Google Scholar]
  10. McCann P. P., Tardif C., Pegg A. E., Diekema K. The dual action of the non-physiological diamines 1,3 diaminopropane and cadaverine on ornithine decarboxylase of HTC cells. Life Sci. 1980 Jun 9;26(23):2003–2010. doi: 10.1016/0024-3205(80)90633-5. [DOI] [PubMed] [Google Scholar]
  11. Mitchell J. L., Augustine T. A., Wilson J. M. Protein factor which induces conversion between Physarum ornithine decarboxylase forms in vitro. Biochim Biophys Acta. 1981 Jan 15;657(1):257–267. doi: 10.1016/0005-2744(81)90149-2. [DOI] [PubMed] [Google Scholar]
  12. Mitchell J. L., Carter D. D. Physical and kinetic distinction of two ornithine decarboxylase forms in Physarum. Biochim Biophys Acta. 1977 Aug 11;483(2):425–434. doi: 10.1016/0005-2744(77)90070-5. [DOI] [PubMed] [Google Scholar]
  13. Mitchell J. L., Carter D. D., Rybski J. A. Control of ornithine decarboxylase activity in Physarum by polyamines. Eur J Biochem. 1978 Dec;92(2):325–331. doi: 10.1111/j.1432-1033.1978.tb12751.x. [DOI] [PubMed] [Google Scholar]
  14. Mitchell J. L., Kottas G. E. Osmotically-induced modification of ornithine decarboxylase in Physarum. FEBS Lett. 1979 Jun 15;102(2):265–268. doi: 10.1016/0014-5793(79)80015-0. [DOI] [PubMed] [Google Scholar]
  15. Mitchell J. L., Mitchell G. K., Carter D. D. Amine-specificity of the inactivating ornithine decarboxylase modification in Physarum polycephalum. Biochem J. 1982 Sep 1;205(3):551–557. doi: 10.1042/bj2050551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mitchell J. L. Ornithine decarboxylase and the ornithine decarboxylase-modifying protein of Physarum polycephalum. Methods Enzymol. 1983;94:140–146. doi: 10.1016/s0076-6879(83)94022-3. [DOI] [PubMed] [Google Scholar]
  17. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  18. Pegg A. E., Conover C., Wrona A. Effects of aliphatic diamines on rat liver ornithine decarboxylase activity. Biochem J. 1978 Mar 15;170(3):651–660. doi: 10.1042/bj1700651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pritchard M. L., Seely J. E., Pösö H., Jefferson L. S., Pegg A. E. Binding of radioactive alpha-difluoromethylornithine to rat liver ornithine decarboxylase. Biochem Biophys Res Commun. 1981 Jun;100(4):1597–1603. doi: 10.1016/0006-291x(81)90701-4. [DOI] [PubMed] [Google Scholar]
  20. Russell D. H. Posttranslational modification of ornithine decarboxylase by its product putrescine. Biochem Biophys Res Commun. 1981 Apr 30;99(4):1167–1172. doi: 10.1016/0006-291x(81)90741-5. [DOI] [PubMed] [Google Scholar]
  21. Scheinberg D. A., Strand M. A brain membrane protein similar to the rat src gene product. Proc Natl Acad Sci U S A. 1981 Jan;78(1):55–59. doi: 10.1073/pnas.78.1.55. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Seely J. E., Pösö H., Pegg A. E. Purification of ornithine decarboxylase from kidneys of androgen-treated mice. Biochemistry. 1982 Jul 6;21(14):3394–3399. doi: 10.1021/bi00257a023. [DOI] [PubMed] [Google Scholar]
  23. Tyagi A. K., Tabor H., Tabor C. W. Inactivation of yeast ornithine decarboxylase by polyamines in vivo does not result from the incorporation of polyamines into enzyme protein. Biochem Biophys Res Commun. 1982 Nov 30;109(2):533–540. doi: 10.1016/0006-291x(82)91754-5. [DOI] [PubMed] [Google Scholar]

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