Abstract
Polyamine oxidases, defined in this chapter as amine oxidases capable of catalyzing the oxidation of spermidine or spermine (1,2) (see Chapter 1), are present in plants, bacteria, fungi, protozoa, worms, and all mammalian cells (2,3) (see references in Chapter 1). In mammalian cells spermine and spermidine can be converted back to putrescine by the pathways shown in Chapter 1, Fig. 5. The first step is the acetylation of an aminopropyl group to give the N 1-acetyl derivative. This is cleaved by polyamine oxidase to form an aldehyde, 3-acetamidopropanal and either spermidine or putrescine (see Chapter 1, Fig. 6). However, acetylation is not a prerequisite for oxidation, and nonacetylated spermidine or spermine can also act as substrates, but less efficiently. Oxidation of spermine or spermidine by some tissue enzymes is stimulated if benzaldehyde is added to the assay mixture (Table 1). This stimulation results in a lowering of the K m value for the substrate, probably as a consequence of Schiff base formation between the aldehyde and the primary amino groups of spermine or spermidine, and appears to mimic the effects of in vivo acetylation. Use of benzaldehyde in this assay substitutes for the nonavailability of radiolabeled N 1-acetylspermine or N 1-acetylspermidine, the preferred substrates of tissue polyamine oxidase. The effect of benzaldehyde addition on the particular polyamine oxidase being assayed should always be examined before routinely including it in the assay mixture.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Morgan, D. M. L. (1985) Polyamine oxidases. Biochem. Soc. Trans. 13, 322–326.
Morgan, D. M. L. (1989) Polyamine oxidase and oxidised polyamines, in Physiology of the Polyamines, vol. I (Bachrach, U. and Heimer, Y., eds.), CRC, Boca Raton, FL, pp. 203–229.
Müller, S. and Walter, R. D. (1992) Purification and characterization of polyamine oxidase from Ascaris suum. Biochem. J. 283, 75–80.
Gahl, W. A., Vale, A. M., and Pitot, H. C. (1982) Spermidine oxidase in human pregnancy serum. Probable identity with diamine oxidase. Biochem. J. 201, 161–164.
Morgan, D. M. L. (1983) Polyamine oxidation and human pregnancy. Adv. Polyamine Res. 4, 155–167.
Morgan, D. M. L. (1985) Human pregnancy-associated polyamine oxidase: partial purification and properties. Biochem. Soc. Trans. 13, 351, 352.
Morgan, D. M. L. and Ilei, G. (1981) Radiochemical estimation of serum polyamine oxidase activity in human pregnancy. Med. Lab. Sci. 38, 49–56.
Ferrante, A., Storer, R. J., and Cleland, L. J. (1990) Polyamine oxidase activity in rheumatoid arthritis synovial fluid. Clin. Exper. Immunol. 80, 373–375.
Inoue, H. and Mizutani, A. (1973) A new method for isolation of polyamines from animal tissues. Anal. Biochem. 56, 408–416.
Lowry, O. H., Rosenbrough, N. J., Farr, A. L., and Randall, R. J. (1951) Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275.
Bradford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254.
Höltta, E. (1983) Polyamine oxidase (rat liver). Methods Enzymol. 94, 306–311.
Holtta, E. (1977) Oxidation of spermidine and spermine in rat liver: purification and properties of polyamine oxidase. Biochemistry 16, 91–100.
Bolkenius, F. N., and Seiler, N. (1981) Acetylderivatives as intermediates in polyamine catabolism. Int. J. Biochem. 13, 287–292.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1998 Humana Press Inc.
About this protocol
Cite this protocol
Morgan, D.M.L. (1998). Radiochemical Estimation of Polyamine Oxidase. In: Morgan, D.M.L. (eds) Polyamine Protocols. Methods in Molecular Biology™, vol 79. Humana Press. https://doi.org/10.1385/0-89603-448-8:97
Download citation
DOI: https://doi.org/10.1385/0-89603-448-8:97
Publisher Name: Humana Press
Print ISBN: 978-0-89603-448-8
Online ISBN: 978-1-59259-565-5
eBook Packages: Springer Protocols