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Properties of recombinant human N1-acetylpolyamine oxidase (hPAO): potential role in determining drug sensitivity

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Abstract

The recent cloning of the mammalian gene coding for N1-acetylpolyamine oxidase (PAO) provides the opportunity to directly examine the role of human PAO (hPAO) in polyamine homeostasis as well as its potential role in determining cellular response to antitumor polyamine analogues. To facilitate the study of this enzyme, the production, purification, and characterization of the recombinant hPAO is reported. hPAO oxidizes N1-acetylspermidine (Km=2.1 μM, Kcat=15.0 s−1) and has very high affinity for N1-acetylspermine (Km=0.85 μM, Kcat=31.7 s−1). The recombinant hPAO does not efficiently oxidize spermine, thereby demonstrating a significant difference in substrate specificity from the previously described human spermine oxidase PAOh1/SMO. Importantly, hPAO demonstrates the ability to oxidize a subset of antitumor polyamine analogues, suggesting that this oxidase activity could have a significant effect on determining tumor sensitivity to these or similar agents. Transfection of A549 human lung cancer cells with an hPAO-expressing plasmid leads to a profound decrease in sensitivity to those analogues which act as substrates, confirming its potential to alter drug response. One similarity that hPAO shares with human PAOh1/SMO, is that certain oligoamine analogues are potent inhibitors of its oxidase activity. The results of these studies demonstrate how changes in polyamine catabolism may affect drug response.

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Abbreviations

hPAO:

Human N1-acetylpolyamine oxidase

PAOh1/SMO:

Human spermine oxidase

SSAT:

Spermidine/spermine N1-acetyltransferase

BENSpm:

N1,N11-bis(ethyl)norspermine

CPENSpm:

N1-ethyl-N11-(cyclopropyl)methyl-4,8-diazaundecane

CHENSpm:

N1-ethyl-N11-(cycloheptyl)methyl-4,8-diazaundecane

IPENSpm:

(S)-N1-(2-methyl-1-butyl)-N11-ethyl-4,8-diazaundecane

MDL 72,527:

N1,N4-bis(2,3-butadienyl)-1,4-butanediamine

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Acknowledgements

This research was supported by National Institutes of Health grants CA51085, CA85509, CA88843, and CA98454.

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Authors and Affiliations

  1. The Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Bunting Blaustein Building, 1650 Orleans Street, Baltimore, MD, 21231, USA

    Yanlin Wang, Amy Hacker, Tracy Murray-Stewart, Alison V. Fraser & Robert A. Casero Jr

  2. SLIL Biomedical Corporation, Madison, WI, 53711, USA

    Benjamin Frydman & Aldonia Valasinas

  3. Department of Pharmaceutical Sciences, Wayne State University, 539 Shapero Hall, Detroit, MI, 48202, USA

    Patrick M. Woster

Authors
  1. Yanlin Wang
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  2. Amy Hacker
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  4. Benjamin Frydman
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  6. Alison V. Fraser
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  7. Patrick M. Woster
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  8. Robert A. Casero Jr
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Correspondence to Robert A. Casero Jr.

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Dedicated to Ben Frydman who died on 25 September 2004.

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Wang, Y., Hacker, A., Murray-Stewart, T. et al. Properties of recombinant human N1-acetylpolyamine oxidase (hPAO): potential role in determining drug sensitivity. Cancer Chemother Pharmacol 56, 83–90 (2005). https://doi.org/10.1007/s00280-004-0936-5

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  • DOI: https://doi.org/10.1007/s00280-004-0936-5

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