Newly synthesized proteins and lipids are transported across the Golgi complex via different mechanisms whose respective roles are not completely clear. We previously identified a non-vesicular intra-Golgi transport pathway for glucosylceramide (GlcCer)—the common precursor of the different series of glycosphingolipids—that is operated by the cytosolic GlcCer-transfer protein FAPP2 (also known as PLEKHA8) (ref. 1). However, the molecular determinants of the FAPP2-mediated transfer of GlcCer from the cis-Golgi to the trans-Golgi network, as well as the physiological relevance of maintaining two parallel transport pathways of GlcCer—vesicular and non-vesicular—through the Golgi, remain poorly defined. Here, using mouse and cell models, we clarify the molecular mechanisms underlying the intra-Golgi vectorial transfer of GlcCer by FAPP2 and show that GlcCer is channelled by vesicular and non-vesicular transport to two topologically distinct glycosylation tracks in the Golgi cisternae and the trans-Golgi network, respectively. Our results indicate that the transport modality across the Golgi complex is a key determinant for the glycosylation pattern of a cargo and establish a new paradigm for the branching of the glycosphingolipid synthetic pathway.

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We thank A. Luini, C. Wilson and D. Priestman for discussions, A. Egorova for help with electron microscopy, G. Liebisch, A. Sigruener and G. Schmitz for lipidomic analysis. M.A.D.M. acknowledges the support of Telethon (GSP08002 and GGP06166), Associazione Italiana per la Ricerca sul Cancro (AIRC) (IG 8623), and the EU (FP7 Lipidomicnet). G.D.’A. acknowledges the support of AIRC (MFAG 10585). P.M. acknowledges the support of Academy of Finland and Sigrid Jusélius Foundation. C.-C.C. was funded by a Study Abroad Scholarship from the Taiwan Ministry of Education.

Author information


  1. Telethon Institute of Genetics and Medicine, Via Pietro Castellino 111, 80131 Naples, Italy

    • Giovanni D’Angelo
    • , Elena Polishchuk
    • , Michele Santoro
    • , Fabrizio Capuani
    •  & Maria Antonietta De Matteis
  2. Institute of Protein Biochemistry National Research Council, Via Pietro Castellino 111, 80131 Naples, Italy

    • Giovanni D’Angelo
    • , Antonio Varriale
    •  & Sabato D’Auria
  3. Laboratory of Molecular Traffic, Department of Molecular and Cellular Biology, Institute for Molecular and Cellular Regulation, Gunma University, Gunma 371-8512, Japan

    • Takefumi Uemura
    • , Takashi Sato
    •  & Akihiro Harada
  4. Department of Pharmacology, University of Oxford, Mansfield Road, Oxford OX1 3QT, UK

    • Chia-Chen Chuang
    •  & Frances M. Platt
  5. Department of Biosciences, Biochemistry, Åbo Akademi University, Artillerigatan 6 A III, BioCity, FI-20520 Turku, Finland

    • Henna Ohvo-Rekilä
    •  & Peter Mattjus
  6. Department of Cell Biology and Oncology, Consorzio Mario Negri Sud, Via Nazionale 8/A, 66030 Santa Maria Imbaro, Chieti, Italy

    • Giuseppe Di Tullio
    •  & Tiziana Daniele
  7. Institut Curie Centre de Recherche, 26 rue d’Ulm, 75248 Paris Cedex 05, France

    • Ludger Johannes
  8. CNRS, UMR144, F-75248 Paris, France

    • Ludger Johannes
  9. Dipartimento di Scienze Chimiche and CEINGE Biotecnologie Avanzate, Università di Napoli FedericoII, ViaGaetano Salvatore 482, 80145 Napoli, Italy

    • Maria Monti
    •  & Piero Pucci
  10. MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH, UK

    • Roger L. Williams
    •  & John E. Burke
  11. Department of Cell Biology, Osaka University, Osaka 565-0871, Japan

    • Akihiro Harada


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M.A.D.M. supervised the entire project; M.A.D.M. and G.D.’A. wrote the manuscript with comments from all co-authors; G.D.’A., with the help of M.S., designed and conducted at TIGEM the experiments of sphingolipid labelling, membrane trafficking, immuno-localization and controlled proteolysis. M.S. designed the strategy and produced plasmid vectors. M.S. and G.D.T. prepared recombinant proteins, anti-FAPP2 and anti-BET3 antibodies. T.U. and T.S. generated and characterized FAPP2geo/geo and FAPP2−/− mice under the supervision of A.H. C.-C.C. conducted the HPLC measurements of GSLs under the supervision of F.M.P. L.J. provided the Cy3-ShTxB. E.P. and T.D. conducted the electron microscopy experiments. H.O.-R. conducted the surface plasmon resonance experiments under the supervision of P.M. A.V. conducted the tryptophan fluorescence and circular dichroism experiments under the supervision of S.D’.A. F.C. and G.D’.A. produced the mathematical model for GSL metabolism. M.M. and P.P. performed and interpreted the MS analysis; R.L.W. and J.E.B. performed and interpreted the HDX analysis.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Maria Antonietta De Matteis.

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