Cellular and Molecular Life Sciences

, Volume 71, Issue 22, pp 4457–4470 | Cite as

Processing of heparanase is mediated by syndecan-1 cytoplasmic domain and involves syntenin and α-actinin

Research Article
First Online:
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Abstract

Heparanase activity plays a decisive role in cell dissemination associated with cancer metastasis. Cellular uptake of heparanase is considered a pre-requisite for the delivery of latent 65-kDa heparanase to lysosomes and its subsequent proteolytic processing and activation into 8- and 50-kDa protein subunits by cathepsin L. Heparan sulfate proteoglycans, and particularly syndecan, are instrumental for heparanase uptake and activation, through a process that has been shown to occur independent of rafts. Nevertheless, the molecular mechanism underlying syndecan-mediated internalization outside of rafts is unclear. Here, we examined the role of syndecan-1 cytoplasmic domain in heparanase processing, utilizing deletion constructs lacking the entire cytoplasmic domain (Delta), the conserved (C1 or C2), or variable (V) regions. Heparanase processing was markedly increased following syndecan-1 over-expression; in contrast, heparanase was retained at the cell membrane and its processing was impaired in cells over-expressing syndecan-1 deleted for the entire cytoplasmic tail. We have next revealed that conserved domain 2 (C2) and variable (V) regions of syndecan-1 cytoplasmic tail mediate heparanase processing. Furthermore, we found that syntenin, known to interact with syndecan C2 domain, and α actinin are essential for heparanase processing.

Keywords

Heparanase Uptake Syndecan Cytoplasmic tail Processing 
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Notes

Acknowledgments

This work was supported by grants from the Israel Science Foundation (Grant 593/10); National Cancer Institute, NIH (Grant CA106456); the Israel Cancer Research Fund (ICRF); the German-Israeli Foundation for Scientific Research and Development (GIF), and the Rappaport Family Institute Fund to I. Vlodavsky. I. Vlodavsky is a Research Professor of the ICRF.

Supplementary material

18_2014_1629_MOESM1_ESM.tif (1.4 mb)
U87 glioma cells over-expressing the syndecan-1 variants were plated on glass coverslips for 24 h. Cells were then fixed and subjected to immunofluorescent staining applying anti-vinculin antibody (A, lower panels, green) and TRITC-phalloidin (A, upper panels, red). The number of vinculin-positive focal contacts, counted in at least 12 cells, is shown graphically in panel (B). Note increased vinculin staining in cells over-expressing wild-type syndecan-1 and decreased staining following deletion of the entire cytoplasmic tail (Delta) or the V region (V). p = 0.001, 0.01, and 0.05 for Mock vs. WT, Mock vs. V, and Mock vs. Delta, respectively (TIFF 1426 kb)

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Copyright information

© Springer Basel 2014

Authors and Affiliations

  1. 1.Cancer and Vascular Biology Research Center, the Bruce Rappaport Faculty of MedicineTechnionHaifaIsrael

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