The degradation (by distinct pathways) of human d-amino acid oxidase and its interacting partner pLG72 – two key proteins in d-serine catabolism in the brain

Authors

  • Pamela Cappelletti,

    1. Dipartimento di Biotecnologie e Scienze della Vita, Università degli Studi dell'Insubria, Varese, Italy
    2. Centro Interuniversitario di Ricerca in Biotecnologie Proteiche ‘The Protein Factory’, Politecnico di Milano, ICRM-CNR Milano and Università degli studi dell'Insubria, Varese, Italy
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  • Paola Campomenosi,

    1. Dipartimento di Biotecnologie e Scienze della Vita, Università degli Studi dell'Insubria, Varese, Italy
    2. Centro Interuniversitario di Ricerca in Biotecnologie Proteiche ‘The Protein Factory’, Politecnico di Milano, ICRM-CNR Milano and Università degli studi dell'Insubria, Varese, Italy
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  • Loredano Pollegioni,

    Corresponding author
    1. Dipartimento di Biotecnologie e Scienze della Vita, Università degli Studi dell'Insubria, Varese, Italy
    2. Centro Interuniversitario di Ricerca in Biotecnologie Proteiche ‘The Protein Factory’, Politecnico di Milano, ICRM-CNR Milano and Università degli studi dell'Insubria, Varese, Italy
    • Correspondence

      L. Pollegioni, Dipartimento di Biotecnologie e Scienze della Vita, Università degli Studi dell'Insubria, via J.H. Dunant 3, Varese 21100, Italy

      Fax: +39 0332 421500

      Tel: +39 0332 421506

      E-mail: loredano.pollegioni@uninsubria.it

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  • Silvia Sacchi

    1. Dipartimento di Biotecnologie e Scienze della Vita, Università degli Studi dell'Insubria, Varese, Italy
    2. Centro Interuniversitario di Ricerca in Biotecnologie Proteiche ‘The Protein Factory’, Politecnico di Milano, ICRM-CNR Milano and Università degli studi dell'Insubria, Varese, Italy
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Abstract

Human d-amino acid oxidase (EC 1.4.3.3; hDAAO) is a peroxisomal flavoenzyme significantly enriched in the mammalian brain. hDAAO has been proposed to play (with serine racemase; EC 5.1.1.18) an essential role in the catabolism of d-serine, an ‘atypical’ key signalling molecule that acts as allosteric activator of the N-methyl-d-aspartate-type glutamate receptor (NMDAr). hDAAO and its interacting partner pLG72 have been related to schizophrenia, a highly disabling psychiatric disorder in which a dysfunction of NMDA-mediated neurotransmission is widely assumed to occur. We previously demonstrated that the d-serine cellular concentration depends on hDAAO and pLG72 expression levels and that newly-synthesized hDAAO interacts with its modulator in the cytosol, being progressively destabilized and inactivated. To obtain insight into the mechanisms regulating cellular d-serine levels, we investigated the degradation pathways of hDAAO and pLG72 in U87 glioblastoma cells stably expressing enhanced yellow fluorescent protein-hDAAO (peroxisomal), hDAAO-enhanced yellow fluorescent protein (cytosolic) or pLG72-enhanced cyan fluorescent protein (mitochondrial) proteins. hDAAO is a long-lived protein: the peroxisomal fraction of this flavoprotein is degraded via the lysosomal/endosomal pathway (and blocking this pathway increases the cellular hDAAO activity and decreases d-serine levels), whereas the cytosolic portion is ubiquitinated and targeted to the proteasome. By contrast, pLG72 shows a rapid turnover (t1/2 ≈ 25–40 min) and is degraded via the proteasome system, albeit not ubiquitinated. Overexpression of pLG72 increases the turnover of hDAAO, in turn playing a protective role against excessive d-serine depletion.

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