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New insights into the operative network of FaEO, an enone oxidoreductase from Fragaria x ananassa Duch.

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Abstract

The 2-methylene-furan-3-one reductase or Fragaria x ananassa Enone Oxidoreductase (FaEO) catalyses the last reductive step in the biosynthesis of 4-hydroxy-2,5-dimethyl-3(2H)-furanone, a major component in the characteristic flavour of strawberries. In the present work, we describe the association between FaEO and the vacuolar membrane of strawberry fruits. Even if FaEO lacks epitopes for stable or transient membrane-interactions, it contains a calmodulin-binding region, suggesting that in vivo FaEO may be associated with the membrane via a peripheral protein complex with calmodulin. Moreover, we also found that FaEO occurs in dimeric form in vivo and, as frequently observed for calmodulin-regulated proteins, it may be expressed in different isoforms by alternative gene splicing. Further mass spectrometry analysis confirmed that the isolated FaEO consists in the already known isoform and that it is the most characteristic during ripening. Finally, a characterization by absorption spectroscopy showed that FaEO has specific flavoprotein features. The relevance of these findings and their possible physiological implications are discussed.

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Acknowledgements

This work was carried out with support from the Marie Curie program ‘‘European Reintegration Grant’’ (PERG05-GA-2009-247789) and the program ‘‘FSE SARDEGNA 2007–2013, Legge Regionale 7 agosto 2007, n. 7, Promozione della ricerca scientifica e dell’innovazione tecnologica in Sardegna’’.

Author contributions

DP conceived the study, participated in its design and coordination, carried out the membranes preparation, participated in the biochemical studies and drafted the manuscript. GC participated in the design of the study and in the membranes preparation. DF partecipate in the bioinformatic studies, participated in the biochemical studies and helped to draft the manuscript. JK carried out the mass spectrometry analysis and helped to draft the manuscript. FE and FP participated in the biochemical studies.

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    1. Laboratory of Plant Physiology and Photobiology, Department of Life and Environmental Sciences, University of Cagliari, V.le S. Ignazio da Laconi 13, 09123, Cagliari, Italy

      Gabriella Collu, Domenica Farci & Dario Piano

    2. Department of Molecular Sensory Systems, Center of Advanced European Studies and Research (caesar), Ludwig-Erhard-Allee 2, 52175, Bonn, Germany

      Domenica Farci

    3. Department of Life and Environmental Sciences, University of Cagliari, Cittadella Universitaria, 09042, Monserrato, Italy

      Francesca Esposito & Francesca Pintus

    4. Leibniz Institute on Aging – Fritz Lipmann Institute, Beutebergstraβe 11, 07745, Jena, Germany

      Joanna Kirkpatrick

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    1. Gabriella Collu
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    Correspondence to Dario Piano.

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    Gabriella Collu and Domenica Farci have contributed equally to the work.

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    Collu, G., Farci, D., Esposito, F. et al. New insights into the operative network of FaEO, an enone oxidoreductase from Fragaria x ananassa Duch.. Plant Mol Biol 94, 125–136 (2017). https://doi.org/10.1007/s11103-017-0597-5

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