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Measurement of Spermidine/Spermine N 1-Acetyltransferase Activity

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Book cover Polyamine Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 79))

Abstract

The breakdown of the polyamines, spermidine and spermine (Fig. 1), is a two-step process involving in the first reaction, spermidine/spermine N 1-acetyltransferase (abbreviated to N 1-SAT or SSAT), and in the second reaction, polyamine oxidase (PAO). N 1-SAT is the rate-limiting enzyme in this catabolic pathway. It is a highly inducible enzyme whose activity is increased by a range of hormones and drugs (13) N 1-SAT acetylates primary amino groups separated from another nitrogen atom by a three-carbon aliphatic chain (4). The reaction appears to occur by an ordered Bi Bi mechanism. The enzyme acetylates both spermine and spermidine, but it is unclear which of these polyamines is the preferred substrate in vivo.

The polyamine catabolic pathway.

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References

  1. Matsui, I. and Pegg, A. E. (1980) Increase in acetylation of spermidine in rat liver extracts brought about by treatment with carbon tetrachloride. Biochem. Biophys. Res. Commun. 92, 1009–1015.

    Article  PubMed  CAS  Google Scholar 

  2. Casero, R. A. and Pegg, A. E. (1993) Spermidine/spermine N1-acetyltransferase—the turning point in polyamine metabolism. FASEB J. 7, 653–661.

    PubMed  CAS  Google Scholar 

  3. Wallace, H. M., Nuttall, M. E., and Robinson, F. C. (1988) Acetylation of spermidine and methylglyoxal bis(guanylhydrazone) in baby hamster kidney cells (BHK-21/C13). Biochem. J. 253, 223–227.

    PubMed  CAS  Google Scholar 

  4. Della-Ragione, F. and Pegg, A. E. (1983) Studies on the specificity and kinetics of rat liver spermidine/spermine N1-acetyltransferase. Biochem. J. 213, 701–706.

    PubMed  CAS  Google Scholar 

  5. Seiler, N. (1987) Functions of polyamine acetylation. Can. J. Physiol. Pharmacol. 65, 2024–2035.

    PubMed  CAS  Google Scholar 

  6. Wallace, H. M. and Ken, H. M. (1981) Uptake and excretion of polyamines from baby hamster kidney cells (BHK-21/Cl3): the effect of serum on confluent cell cultures. Biochim. Biophys. Acta 676, 25–30.

    PubMed  CAS  Google Scholar 

  7. Wallace, H. M. (1987) Polyamine catabolism in mammalian cells excretion and acetylation. Med. Sci. Res. 15, 1437–1440.

    CAS  Google Scholar 

  8. Pegg, A. E., Pakala, R., and Bergeron, R. J. (1990) Induction of spermidine/spermine N1-acetyltransferase activity in Chinese hamster ovary cells by N1, N12 bis(ethyl)norspermidine and related compounds. Biochem. J. 267, 331–338.

    PubMed  CAS  Google Scholar 

  9. Melvin, M. A. L. and Ken, H. M. (1978) Polyamine metabolism in BHK-21/Cl3 cells. Loss of spermidine from cells following transfer to serum-depleted medium. Exper. Cell Res. 111, 231–236.

    Article  CAS  Google Scholar 

  10. Mackarel, A. J. and Wallace H. M. (1992) Effect of 5-fluorouracil on polyamine excretion in human colonic cancer cells. Biochem. Soc. Trans. 21, 50S.

    Google Scholar 

  11. Brunton, V. G., Grant, M. H., and Wallace, H. M. (1990) Spermine toxicity and glutathione depletion in BHK-21/Cl3 cells. Biochem. Pharmacol. 40, 1893–1900.

    Article  PubMed  CAS  Google Scholar 

  12. Abdel-Monen, M. M., Ohno, K., Fortuny, I. E., and Theologides, A. (1975) Acetylspermidines in human urine. Lancet ii, 1210.

    Article  Google Scholar 

  13. Loser, C., Folsch, U. R., Papronty, C., and Creutzfeld, W. (1990) Polyamines in colorectal cancer. Evaluation of polyamine concentrations in colon tissue, serum and urine of 50 patients with colorectal cancer. Cancer 65, 958–966.

    Article  PubMed  CAS  Google Scholar 

  14. Russell, D. H., Levy, C. C., Schimpff, S. C., and Hawk, I. A. (1971) Urinary polyamines in cancer patients. Cancer Res. 31, 1555–1558.

    PubMed  CAS  Google Scholar 

  15. Lipton, A., Sheehan, L. M., and Kessler, G. F. (1975) Urinary polyamine levels in human cancer. Cancer 35, 464–468.

    Article  PubMed  CAS  Google Scholar 

  16. Seiler, N., Bolkenius, F. N., and Knodgen, B. (1985) The influence of catabolic reactions on polyamine excretion. Biochem. J. 225, 219–226.

    PubMed  CAS  Google Scholar 

  17. Poulin, R., Wechter, R. S., and Pegg, A. E. (1991) An early enlargement of the putrescine pool is required for growth of L1210 mouse leukaemia cells under hypoosmotic stress. J. Biol. Chem. 266, 6142–6151.

    PubMed  CAS  Google Scholar 

  18. Poulin, R., Coward, J. K., Lakanen, L. R., and Pegg, A. E. (1993) Enhancement of the spermidine uptake system and lethal effects of spermidine overaccumulation in ornithine decarboxylase overproducing L1210 cells under hypoosmotic stress. J. Biol. Chem. 268, 4690–4698.

    PubMed  CAS  Google Scholar 

  19. Libby, P. R. (1978) Calf liver nuclear N-acetyltransferases. Purification and properties of two enzymes with both spermidine acetyltransferase and histone acetyltransferase activities. J. Biol. Chem. 253, 233–237.

    PubMed  CAS  Google Scholar 

  20. Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. (1951) Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275.

    PubMed  CAS  Google Scholar 

  21. Seiler, N. and Knodgen, B. (1980) Hugh performance liquid chromatographic procedure for the simultaneous determination of the natural polyamines and then monoacetyl derivatives. J. Chromatog. 221, 227–235.

    Article  CAS  Google Scholar 

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© 1998 Humana Press Inc.

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Wallace, H.M., Evans, D.M. (1998). Measurement of Spermidine/Spermine N 1-Acetyltransferase Activity. In: Morgan, D.M.L. (eds) Polyamine Protocols. Methods in Molecular Biology™, vol 79. Humana Press. https://doi.org/10.1385/0-89603-448-8:59

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  • DOI: https://doi.org/10.1385/0-89603-448-8:59

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-448-8

  • Online ISBN: 978-1-59259-565-5

  • eBook Packages: Springer Protocols

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