Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1997 Feb 15;322(Pt 1):297–302. doi: 10.1042/bj3220297

CGP 48664, a potent and specific S-adenosylmethionine decarboxylase inhibitor: effects on regulation and stability of the enzyme.

F Svensson 1, H Mett 1, L Persson 1
PMCID: PMC1218191  PMID: 9078276

Abstract

Mammalian S-adenosylmethionine decarboxylase (AdoMetDC) catalyses a regulatory important step in the biosynthesis of polyamines and is a potential target for therapeutic agents against various parasitic diseases and proliferative disorders. In the present study we examined the effects of a newly synthesized AdoMetDC inhibitor. 4-amidinoindan-1-one 2'-amidinohydrazone (CGP 48664), on polyamine metabolism in the mouse leukaemia cell line L1210. Treatment of the cells with 2 microM CGP 48664 led to a depletion of cellular spermidine and spermine. The putrescine content, in contrast, was markedly increased. Cells seeded in the presence of the inhibitor showed a significant decrease in growth rate, which was fully reversed by the addition of 2 microM spermidine or 1 microM spermine. The syntheses of ornithine decarboxylase and AdoMetDC were greatly increased in cells treated with CGP 48664. These increases were not correlated with similar changes in the mRNA levels, indicating the involvement of a translational mechanism. CGP 48664 was demonstrated to be a very poor competitor of spermidine uptake in the L1210 cells. L1210 cells deficient in polyamine transport were as sensitive to the antiproliferative effect of the inhibitor as were the parental cells, indicating that CGP 48664 did not enter the cells by the polyamine transport system. In addition to inhibiting AdoMetDC, CGP 48664 stabilized the enzyme against degradation. In the present study we also demonstrated that aminoguanidine (AMG), which is frequently used in cellular systems to inhibit any action of serum polyamine oxidase, apparently inhibits AdoMetDC by an irreversible mechanism that markedly stabilizes the enzyme against proteolytic degradation. CGP 48664 and the parental compound methylglyoxal bis(guanylhydrazone), which is also a potent inhibitor of AdoMetDC, contain one or two AMG-like moieties; the importance of these residues in the inhibition of AdoMetDC is discussed.

Full Text

The Full Text of this article is available as a PDF (336.3 KB).

Selected References

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

  1. Ask A., Persson L., Heby O. Increased survival of L1210 leukemic mice by prevention of the utilization of extracellular polyamines. Studies using a polyamine-uptake mutant, antibiotics and a polyamine-deficient diet. Cancer Lett. 1992 Sep 14;66(1):29–34. doi: 10.1016/0304-3835(92)90276-2. [DOI] [PubMed] [Google Scholar]
  2. Byers T. L., Ganem B., Pegg A. E. Cytostasis induced in L1210 murine leukaemia cells by the S-adenosyl-L-methionine decarboxylase inhibitor 5'-([(Z)-4-amino-2-butenyl]methylamino)-5'-deoxyadenosine may be due to hypusine depletion. Biochem J. 1992 Nov 1;287(Pt 3):717–724. doi: 10.1042/bj2870717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Byers T. L., Wiest L., Wechter R. S., Pegg A. E. Effects of chronic 5'-([(Z)-4-amino-2-butenyl]methylamino)-5'-deoxy- adenosine (AbeAdo) treatment on polyamine and eIF-5A metabolism in AbeAdo-sensitive and -resistant L1210 murine leukaemia cells. Biochem J. 1993 Feb 15;290(Pt 1):115–121. doi: 10.1042/bj2900115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Casero R. A., Jr, Pegg A. E. Spermidine/spermine N1-acetyltransferase--the turning point in polyamine metabolism. FASEB J. 1993 May;7(8):653–661. [PubMed] [Google Scholar]
  5. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  6. Demetriou A. A., Cohn M. S., Tabor C. W., Tabor H. Identification of pyruvate in S-adenosylmethionine decarboxylase from rat liver. J Biol Chem. 1978 Mar 10;253(5):1684–1686. [PubMed] [Google Scholar]
  7. Grens A., Steglich C., Pilz R., Scheffler I. E. Nucleotide sequence of the Chinese hamster ornithine decarboxylase gene. Nucleic Acids Res. 1989 Dec 25;17(24):10497–10497. doi: 10.1093/nar/17.24.10497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Heby O., Persson L. Molecular genetics of polyamine synthesis in eukaryotic cells. Trends Biochem Sci. 1990 Apr;15(4):153–158. doi: 10.1016/0968-0004(90)90216-x. [DOI] [PubMed] [Google Scholar]
  9. Holm I., Persson L., Pegg A. E., Heby O. Effects of S-adenosyl-1,8-diamino-3-thio-octane and S-methyl-5'-methylthioadenosine on polyamine synthesis in Ehrlich ascites-tumour cells. Biochem J. 1989 Jul 1;261(1):205–210. doi: 10.1042/bj2610205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Joe Y. A., Park M. H. Structural features of the eIF-5A precursor required for posttranslational synthesis of deoxyhypusine. J Biol Chem. 1994 Oct 14;269(41):25916–25921. [PubMed] [Google Scholar]
  11. Jänne J., Alhonen L., Leinonen P. Polyamines: from molecular biology to clinical applications. Ann Med. 1991 Aug;23(3):241–259. doi: 10.3109/07853899109148056. [DOI] [PubMed] [Google Scholar]
  12. Kameji T., Pegg A. E. Effect of putrescine on the synthesis of S-adenosylmethionine decarboxylase. Biochem J. 1987 Apr 1;243(1):285–288. doi: 10.1042/bj2430285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kramer D. L., Khomutov R. M., Bukin Y. V., Khomutov A. R., Porter C. W. Cellular characterization of a new irreversible inhibitor of S-adenosylmethionine decarboxylase and its use in determining the relative abilities of individual polyamines to sustain growth and viability of L1210 cells. Biochem J. 1989 Apr 15;259(2):325–331. doi: 10.1042/bj2590325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kramer D., Mett H., Evans A., Regenass U., Diegelman P., Porter C. W. Stable amplification of the S-adenosylmethionine decarboxylase gene in Chinese hamster ovary cells. J Biol Chem. 1995 Feb 3;270(5):2124–2132. doi: 10.1074/jbc.270.5.2124. [DOI] [PubMed] [Google Scholar]
  15. Marton L. J., Pegg A. E. Polyamines as targets for therapeutic intervention. Annu Rev Pharmacol Toxicol. 1995;35:55–91. doi: 10.1146/annurev.pa.35.040195.000415. [DOI] [PubMed] [Google Scholar]
  16. McCann P. P., Pegg A. E. Ornithine decarboxylase as an enzyme target for therapy. Pharmacol Ther. 1992;54(2):195–215. doi: 10.1016/0163-7258(92)90032-u. [DOI] [PubMed] [Google Scholar]
  17. Pajunen A., Crozat A., Jänne O. A., Ihalainen R., Laitinen P. H., Stanley B., Madhubala R., Pegg A. E. Structure and regulation of mammalian S-adenosylmethionine decarboxylase. J Biol Chem. 1988 Nov 15;263(32):17040–17049. [PubMed] [Google Scholar]
  18. Pegg A. E., Corti A., Williams-Ashman H. G. Paradoxical enhancement of S-adenosylmethionine decarboxylase in rat tissues following administration of the specific inhibitor methyl glyoxal bis(guanylhydrazone). Biochem Biophys Res Commun. 1973 May 15;52(2):696–701. doi: 10.1016/0006-291x(73)90768-7. [DOI] [PubMed] [Google Scholar]
  19. Pegg A. E., Erwin B. G., Persson L. Induction of spermidine/spermine N1-acetyltransferase by methylglyoxal bis(guanylhydrazone). Biochim Biophys Acta. 1985 Oct 17;842(2-3):111–118. doi: 10.1016/0304-4165(85)90192-8. [DOI] [PubMed] [Google Scholar]
  20. Pegg A. E. Evidence for the presence of pyruvate in rat liver S-adenosylmethionine decarboxylase. FEBS Lett. 1977 Dec 1;84(1):33–36. doi: 10.1016/0014-5793(77)81051-x. [DOI] [PubMed] [Google Scholar]
  21. Pegg A. E., Jones D. B., Secrist J. A., 3rd Effect of inhibitors of S-adenosylmethionine decarboxylase on polyamine content and growth of L1210 cells. Biochemistry. 1988 Mar 8;27(5):1408–1415. doi: 10.1021/bi00405a003. [DOI] [PubMed] [Google Scholar]
  22. Pegg A. E., McCann P. P. S-adenosylmethionine decarboxylase as an enzyme target for therapy. Pharmacol Ther. 1992 Dec;56(3):359–377. doi: 10.1016/0163-7258(92)90025-u. [DOI] [PubMed] [Google Scholar]
  23. Pegg A. E., McGill S. M. Inhibition of diamine oxidase by 1,1-[(methylethanediylidene)-dinitrilo]-bis-(3-aminoguanidine) and 1,1'-[(methylethanediylidene)-dinitrilo]-diguanidine. Biochem Pharmacol. 1978;27(12):1625–1629. doi: 10.1016/0006-2952(78)90170-3. [DOI] [PubMed] [Google Scholar]
  24. Pegg A. E. Polyamine metabolism and its importance in neoplastic growth and a target for chemotherapy. Cancer Res. 1988 Feb 15;48(4):759–774. [PubMed] [Google Scholar]
  25. Pegg A. E., Williams-Ashman H. G. Stimulation of the decarboxylation of S-adenosylmethionine by putrescine in mammalian tissues. Biochem Biophys Res Commun. 1968 Jan 11;30(1):76–82. doi: 10.1016/0006-291x(68)90715-8. [DOI] [PubMed] [Google Scholar]
  26. Persson L. Antibodies to ornithine decarboxylase. Immunochemical cross-reactivity. Acta Chem Scand B. 1982;36(10):685–688. doi: 10.3891/acta.chem.scand.36b-0685. [DOI] [PubMed] [Google Scholar]
  27. Persson L., Stjernborg L., Holm I., Heby O. Polyamine-mediated control of mammalian S-adenosyl-L-methionine decarboxylase expression: effects on the content and translational efficiency of the mRNA. Biochem Biophys Res Commun. 1989 May 15;160(3):1196–1202. doi: 10.1016/s0006-291x(89)80130-5. [DOI] [PubMed] [Google Scholar]
  28. Regenass U., Caravatti G., Mett H., Stanek J., Schneider P., Müller M., Matter A., Vertino P., Porter C. W. New S-adenosylmethionine decarboxylase inhibitors with potent antitumor activity. Cancer Res. 1992 Sep 1;52(17):4712–4718. [PubMed] [Google Scholar]
  29. Regenass U., Mett H., Stanek J., Mueller M., Kramer D., Porter C. W. CGP 48664, a new S-adenosylmethionine decarboxylase inhibitor with broad spectrum antiproliferative and antitumor activity. Cancer Res. 1994 Jun 15;54(12):3210–3217. [PubMed] [Google Scholar]
  30. Stanley B. A., Pegg A. E. Amino acid residues necessary for putrescine stimulation of human S-adenosylmethionine decarboxylase proenzyme processing and catalytic activity. J Biol Chem. 1991 Oct 5;266(28):18502–18506. [PubMed] [Google Scholar]
  31. Stanley B. A., Pegg A. E., Holm I. Site of pyruvate formation and processing of mammalian S-adenosylmethionine decarboxylase proenzyme. J Biol Chem. 1989 Dec 15;264(35):21073–21079. [PubMed] [Google Scholar]
  32. Stjernborg L., Heby O., Mamont P., Persson L. Polyamine-mediated regulation of S-adenosylmethionine decarboxylase expression in mammalian cells. Studies using 5'-([(Z)-4-amino-2-butenyl]methylamino)-5'-deoxyadenosine, a suicide inhibitor of the enzyme. Eur J Biochem. 1993 Jun 15;214(3):671–676. doi: 10.1111/j.1432-1033.1993.tb17967.x. [DOI] [PubMed] [Google Scholar]
  33. Stjernborg L., Persson L. Stabilization of S-adenosylmethionine decarboxylase by aminoguanidine. Biochem Pharmacol. 1993 Mar 9;45(5):1174–1176. doi: 10.1016/0006-2952(93)90266-y. [DOI] [PubMed] [Google Scholar]
  34. Svensson F., Kockum I., Persson L. Diethylglyoxal bis(guanylhydrazone), a potent inhibitor of mammalian S-adenosylmethionine decarboxylase. Effects on cell proliferation and polyamine metabolism in L1210 leukemia cells. Mol Cell Biochem. 1993 Jul 21;124(2):141–147. doi: 10.1007/BF00929206. [DOI] [PubMed] [Google Scholar]
  35. Svensson F., Persson L. Regulation of ornithine decarboxylase and S-adenosylmethionine decarboxylase in a polyamine auxotrophic cell line. Mol Cell Biochem. 1996 Sep 20;162(2):113–119. doi: 10.1007/BF00227537. [DOI] [PubMed] [Google Scholar]
  36. Tabor C. W., Tabor H. Polyamines. Annu Rev Biochem. 1984;53:749–790. doi: 10.1146/annurev.bi.53.070184.003533. [DOI] [PubMed] [Google Scholar]
  37. Williams-Ashman H. G., Seidenfeld J. Aspects of the biochemical pharmacology of methyl glyoxal bis(guanylhydrazone). Biochem Pharmacol. 1986 Apr 15;35(8):1217–1225. doi: 10.1016/0006-2952(86)90263-7. [DOI] [PubMed] [Google Scholar]
  38. Wu Y. Q., Woster P. M. Irreversible inhibition of human S-adenosylmethionine decarboxylase by the pure diastereomeric forms of S-(5'-deoxy-5'-adenosyl)-1-ammonio-4- methylsulfonio-2-cyclopentene (AdoMac). Biochem Pharmacol. 1995 Apr 18;49(8):1125–1133. doi: 10.1016/0006-2952(95)98510-g. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES