Summary
Putrescine-oxidase activity was found in fetal bovine serum (FBS) with a pH optimum of 8.0 and in adult bovine serum (ABS) with a pH optimum of 9.8. The crude FBS enzyme had a KM for putrescine of 2.58×10−6 m and a Vmax of 0.53 nmol per hr per 50 μl serum. Aminoguanidine competitively inhibited the enzyme with a KI of 1.8×10−8 m. Spermidine and spermine proved competitive inhibitors of putrescine for both the FBS and the crude ABS putrescine oxidases. The Vmax for the ABS putrescine oxidase was 2.10 nmol per hr per 50 μl serum, and the KM for putrescine, 50.3×10−6 m. The K1 of the ABS putrescine oxidase for aminoguanidine was 41×10−6 m. On the basis of both the KM and KI values, the adult serum enzyme, at its optimal pH of 9.8, bound spermidine and spermine more avidly than the smaller putrescine and aminoguanidine; whereas the FBS enzyme, at pH 8.0, bound aminoguanidine and putrescine more tightly than the larger polyamines. Each of the enzymes retained over 80% of its activity after heating at 56°C for 30 min. Applications of these data to the study of polyamines in tissue culture and to the purification of diamine oxidases are discussed.
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References
Cohen, S. S. 1971.Introduction to Polyamines. Prentice Hall, Englewood Cliffs, N.J., pp. 1–179.
Bachrach, U. 1973.Function of Naturally Occurring Polyamines. Academic Press, New York, pp. 1–211.
Zeller, E. A. 1963. Diamine oxidases. In: P. D. Boyer, H. Lardy, and D. Myrback (Eds.),The Enzymes. Vol. 8. Academic Press, London, pp. 313–335.
Pohjanpelto, P., and A. Raina. 1972. Identification of a growth factor produced by human fibroblastsin vitro as putrescine. Nature (London) New Biol. 235: 247–249.
Okuyama, T., and Y. Kobayashi. 1961. Determination of diamine oxidase activity by liquid scintillation counting. Arch. Biochem. Biophys. 95: 242–250.
Dixon, M., and E. C. Webb. 1964.Enzymes. Second Ed. Academic Press, New York, pp. 315–359.
Shore, P. A., and V. H. Cohn, Jr., 1960. Comparative effects of monoamine oxidase inhibitors on monoamine oxidase and diamine oxidase. Biochem. Pharmacol. 5: 91–95.
Heller, J. S., W. F. Fong, and E. S. Canellakis. 1976. Induction of a protein inhibitor to ornithine decarboxylase by the end products of its reaction. Proc. Nat. Acad. Sci. U.S.A. 73: 1858–1862.
Holtta, E. 1977. Oxidation of spermidine and spermine in rat liver: Purification and properties of polyamine oxidase. Biochemistry 16: 91–100.
Janne, J., E. Holtta, P. Haaranen, and K. Elfving. 1973. Polyamines and polyamine-metabolizing enzyme activities in human semen. Clin. Chim. Acta 48: 393–401.
Gahl, W. A., J. E. Changus, and H. C. Pitot. 1976. The effect of spermidine and spermine on proliferationin vitro of fibroblasts from normal and cystic fibrosis patients. Pediatr. Res. 10: 531–535.
Alarcon, R. A., G. E. Foley, and E. J. Modest. 1961. Effects of spermine on mammalian cells. Arch. Biochem. Biophys. 94: 540–541.
Gahl, W. A., and H. C. Pitot. 1978. Reversal by amino-guanidine of the inhibition of proliferation of human fibroblasts by spermidine and spermine. Chem. Biol. Interactions 22: 91–98.
Tabor, C. W., H. Tabor, and U. Bachrach. 1964. Identification of the aminoaldehydes produced by the oxidation of spermine and spermidine with purified plasma amine oxidase. J. Biol. Chem. 239: 2194–2203.
Bachrach, U., S. Abzug, and A. Bekierkunst. 1967. Cytotoxic effect of oxidized spermine on Ehrlich ascites cells. Biochim. Biophys. Acta 134: 174–181.
Hill, C. M., and W. G. Bardsley. 1975. Histamine and related compounds as substrates of diamine oxidase (histaminase). Biochem. Pharmacol. 24: 253–257.
Blaschko, H., and R. Hawes. 1959. Observations on spermine oxidase of mammalain plasma. J. Physiol. 145: 124–131.
Tabor, C. W., H. Tabor, and S. M. Rosenthal. 1954. Purification of amine oxidase from beef plasma. J. Biol. Chem. 208: 645–661.
Yamada, H., and K. T. Yasunobu. 1962. Monoamine oxidase: Purification, crystallization, and properties of plasma monoamine oxidase. J. Biol. Chem. 237: 1511–1516.
Byrd, W. J., D. M. Jacobs, and M. S. Amoss. 1977. Synthetic polyamines added to cultures containing bovine sera reversibly inhibitin vitro parameters of immunity. Nature 267: 621–623.
Morris, D. R., C. M. Jorstad, and C. E. Seyfried. 1977. Inhibition of the synthesis of polyamines and DNA in activated lymphocytes by a combination of α-methylornithine and methylglyoxal bis(guanylhydrazone). Cancer Res. 37: 3169–3172.
Fillingame, R. H., C. M. Jorstad, and D. R. Morris. 1975. Increased cellular levels of spermidine or spermine are required for optimal DNA synthesis in lymphocytes activated by concanavalin A. Proc. Nat. Acad. Sci. U.S.A. 72: 4042–4045.
Lundgren, D. W., J. M. Hankins, M. M. Ulane, and J. W. Willison. 1977. Putrescine uptake in cystic fibrosis fibroblasts. J. Pediatr. 90: 1034–1035.
Baylin, S. B., and S. Margolis. 1975. Purification of histaminase (diamine oxidase) from human pregnancy plasma by affinity chromatography. Biochim. Biophys. Acta 397: 294–306.
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This work was supported in part by a grant from the Cystic Fibrosis Foundation.
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Gahl, W.A., Pitot, H.C. Putrescine-oxidase activity in adult bovine serum and fetal bovine serum. In Vitro 15, 252–257 (1979). https://doi.org/10.1007/BF02618948
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DOI: https://doi.org/10.1007/BF02618948