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Glycosphingolipids are required for sorting melanosomal proteins in the Golgi complex

Sprong, H., Degroote, S., Claessens, T., Oorschot, V., Westerink, B. H. C., Hirabayashi, Y., Klumperman, J., van der Sluijs, P. & van Meer, G., 29-Oct-2001, In : Journal of Cell Biology. 155, 3, p. 369-379 11 p.

Research output: Contribution to journalArticleAcademicpeer-review

  • H. Sprong
  • S. Degroote
  • T. Claessens
  • V. Oorschot
  • B. H. C. Westerink
  • Y. Hirabayashi
  • J. Klumperman
  • P. van der Sluijs
  • G. van Meer

Although glycosphingolipids are ubiquitously expressed and essential for multicellular organisms, surprisingly little is known about their intracellular functions. To explore the role of glycosphingolipids in membrane transport, we used the glycosphingolipid-deficient GM95 mouse melanoma cell line. We found that GM95 cells do not make melanin pigment because tyrosinase, the first and rate-limiting enzyme in melanin synthesis, was not targeted to melanosomes but accumulated in the Golgi complex. However, tyrosinase-related protein 1 still reached melanosomal structures via the plasma membrane instead of the direct pathway from the Golgi. Delivery of lysosomal enzymes from the Golgi complex to endosomes was normal, suggesting that this pathway is not affected by the absence of glycosphingolipids. Loss of pigmentation was due to tyrosinase mislocalization, since transfection of tyrosinase with an extended transmembrane domain, which bypassed the transport block, restored pigmentation. Transfection of ceramide glucosyltransferase or addition of glucosylsphingosine restored tyrosinase transport and pigmentation. We conclude that protein transport from Golgi to melanosomes via the direct pathway requires glycosphingolipids.

Original languageEnglish
Pages (from-to)369-379
Number of pages11
JournalJournal of Cell Biology
Volume155
Issue number3
Publication statusPublished - 29-Oct-2001

    Keywords

  • glycosphingolipid, protein sorting, melanosome, tyrosinase, TRP-1, TYROSINASE-RELATED PROTEIN-1, MOUSE MELANOMA-CELLS, DI-LEUCINE, MDCK CELLS, CERAMIDE GALACTOSYLTRANSFERASE, CYTOPLASMIC TAIL, EPITHELIAL-CELLS, MEMBRANE RAFTS, AP-3 ADAPTER, TRANSPORT

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