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Cell-specific ATP7A transport sustains copper-dependent tyrosinase activity in melanosomes

Nature volume 454, pages 1142–1146 (2008)Cite this article

Abstract

Copper is a cofactor for many cellular enzymes and transporters1. It can be loaded onto secreted and endomembrane cuproproteins by translocation from the cytosol into membrane-bound organelles by ATP7A or ATP7B transporters, the genes for which are mutated in the copper imbalance syndromes Menkes disease and Wilson disease, respectively2. Endomembrane cuproproteins are thought to incorporate copper stably on transit through the trans-Golgi network, in which ATP7A accumulates3 by dynamic cycling through early endocytic compartments4. Here we show that the pigment-cell-specific cuproenzyme tyrosinase acquires copper only transiently and inefficiently within the trans-Golgi network of mouse melanocytes. To catalyse melanin synthesis, tyrosinase is subsequently reloaded with copper within specialized organelles called melanosomes. Copper is supplied to melanosomes by ATP7A, a cohort of which localizes to melanosomes in a biogenesis of lysosome-related organelles complex-1 (BLOC-1)-dependent manner. These results indicate that cell-type-specific localization of a metal transporter is required to sustain metallation of an endomembrane cuproenzyme, providing a mechanism for exquisite spatial control of metalloenzyme activity. Moreover, because BLOC-1 subunits are mutated in subtypes of the genetic disease Hermansky–Pudlak syndrome, these results also show that defects in copper transporter localization contribute to hypopigmentation, and hence perhaps other systemic defects, in Hermansky–Pudlak syndrome.

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Figure 1: Copper transporter ATP7A localizes to melanosomes in wild-type melanocytes.
Figure 2: ATP7A is mislocalized to early endosomes in BLOC-1-deficient melanocytes.
Figure 3: Tyrosinase is present but inactive in BLOC-1-deficient melanocytes.
Figure 4: Copper restores in vitro tyrosinase activity in melanosomes of BLOC-1-deficient melanocytes.

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Acknowledgements

We thank J. Gitlin, M. Petris, B. Eipper, S. Lutsenko and A. Peden for reagents; A. Dancis, L. King and C. Burd for comments; and D. Harper for technical assistance. This work was supported by National Institute of Health grants R01 EY015625 and R21 GM078474 (to M.S.M.), CNRS, Institut Curie and Fondation pour la Recherche Médicale (to G.R.), Wellcome Trust program grant 064583 (to E.V.S. and D.C.B.), and postdoctoral fellowship 0625437U from the American Heart Association (to S.R.G.S.).

Author Contributions S.R.G.S. designed and performed most of the experiments, prepared most of the figures, wrote an initial draft of the manuscript and participated in all stages of manuscript revision. D.T. performed all of the electron microscopy analyses and provided valuable insights into data interpretation. E.V.S. and D.C.B. provided the cell lines used in all of the experiments, performed confirmatory experiments and guided others, provided further data not shown in the manuscript and participated in manuscript revision. G.R. oversaw the electron microscopy analyses, prepared electron micrographs for the figures, contributed substantially to experimental design and participated in manuscript revision. M.S.M. oversaw the entire project, designed many of the experiments in collaboration with S.R.G.S., coordinated work among collaborators and participated in all stages of manuscript revision.

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Authors and Affiliations

  1. Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA,

    Subba Rao Gangi Setty & Michael S. Marks

  2. Institut Curie, Centre de Recherche, Paris F-75248, France ,

    Danièle Tenza & Graça Raposo

  3. Centre National de la Recherche Scientifique, UMR 144, Paris F-75248, France ,

    Danièle Tenza & Graça Raposo

  4. Division of Basic Medical Sciences, Centre for Molecular and Metabolic Signalling, St George’s, University of London, London SW17 0RE, UK

    Elena V. Sviderskaya & Dorothy C. Bennett

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  1. Subba Rao Gangi Setty
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Correspondence to Michael S. Marks.

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Setty, S., Tenza, D., Sviderskaya, E. et al. Cell-specific ATP7A transport sustains copper-dependent tyrosinase activity in melanosomes. Nature 454, 1142–1146 (2008). https://doi.org/10.1038/nature07163

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