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Pacific oyster polyamine oxidase: a protein missing link in invertebrate evolution

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Abstract

Polyamine oxidases catalyse the oxidation of polyamines and acetylpolyamines and are responsible for the polyamine interconversion metabolism in animal cells. Polyamine oxidases from yeast can oxidize spermine, N 1-acetylspermine, and N 1-acetylspermidine, while in vertebrates two different enzymes, namely spermine oxidase and acetylpolyamine oxidase, specifically catalyse the oxidation of spermine, and N 1-acetylspermine/N 1-acetylspermidine, respectively. In this work we proved that the specialized vertebrate spermine and acetylpolyamine oxidases have arisen from an ancestor invertebrate polyamine oxidase with lower specificity for polyamine substrates, as demonstrated by the enzymatic activity of the mollusc polyamine oxidase characterized here. This is the first report of an invertebrate polyamine oxidase, the Pacific oyster Crassostrea gigas (CgiPAO), overexpressed as a recombinant protein. This enzyme was biochemically characterized and demonstrated to be able to oxidase both N 1-acetylspermine and spermine, albeit with different efficiency. Circular dichroism analysis gave an estimation of the secondary structure content and modelling of the three-dimensional structure of this protein and docking studies highlighted active site features. The availability of this pluripotent enzyme can have applications in crystallographic studies and pharmaceutical biotechnologies, including anticancer therapy as a source of hydrogen peroxide able to induce cancer cell death.

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Abbreviations

AcSpd:

N 1-acetylspermidine

AcSpm:

N 1-acetylspermine

APAO:

N 1-acetylpolyamine oxidase

CgiPAO:

Crassostrea gigas polyamine oxidase

FAD:

Flavin adenine dinucleotide

FMN:

Flavin mononucleotide

H2O2 :

Hydrogen peroxide

PAO:

Polyamine oxidase

PAs:

Polyamines

Put:

Putrescine

Spd:

Spermidine

Spm:

Spermine

SMO:

Spermine oxidase

ZmPAO:

Maize polyamine oxidase

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Acknowledgments

We wish to thank the University of Roma Tre for financial support and Prof. Marco Oliverio (Sapienza University of Rome, Italy) for useful discussion.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Sciences, University of Roma Tre, 00146, Rome, Italy

    Manuela Cervelli, Fabio Polticelli, Emanuela Angelucci, Elena Di Muzio, Pasquale Stano & Paolo Mariottini

  2. Interuniversity Consortium of Structural and Systems Biology, 00136, Rome, Italy

    Manuela Cervelli & Paolo Mariottini

  3. National Institute of Nuclear Physics, Roma Tre Section, 00146, Rome, Italy

    Fabio Polticelli

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  1. Manuela Cervelli
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  2. Fabio Polticelli
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  3. Emanuela Angelucci
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Correspondence to Manuela Cervelli.

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Handling Editor: S. Beninati.

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Cervelli, M., Polticelli, F., Angelucci, E. et al. Pacific oyster polyamine oxidase: a protein missing link in invertebrate evolution. Amino Acids 47, 949–961 (2015). https://doi.org/10.1007/s00726-015-1924-2

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  • DOI: https://doi.org/10.1007/s00726-015-1924-2

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