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Cross-linking of glycosphingolipids at the plasma membrane: consequences for intracellular signaling and traffic

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Abstract

Glycosphingolipids (GSLs) are predominantly found in the outer leaflet of the plasma membrane, where they play a role in important processes such as cell adhesion, migration and signaling. However, by which mechanisms GSLs regulate these processes remains elusive. In this study, we therefore took advantage of the fact that some GSLs also serve as receptors for certain protein toxins, which rely on receptor binding for internalization and intoxication. Here, we demonstrate that Shiga and cholera toxins, which both possess multivalent GSL-binding capacity, induce dissociation of the cytosolic cPLA2α–AnxA1 complex in HeLa and HMEC-1 cells. The dissociation is mediated through an increase in cytosolic calcium levels and activation of the tyrosine kinase Syk. Ricin, a protein toxin that does not cross-link surface molecules, has no effect on the same complex. Importantly, we find that antibody-mediated cross-linking of Gb3 and GM1, the GSL receptors for Shiga and cholera toxin, respectively, also induces dissociation. These data demonstrate that cross-linking of GSLs at the plasma membrane mediates the intracellular signaling events resulting in dissociation of the complex. After dissociation, cPLA2α and AnxA1 are translocated to intracellular membranes where they are known to function in regulating membrane transport processes. In conclusion, we have characterized a novel mechanism for cell surface-induced initiation of intracellular signaling and transport events.

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Acknowledgments

We thank Anne Engen for excellent assistance with cell culturing, Anne-Mari Gjestvang Pedersen and Roger Simm for purification of Shiga toxins, and Maria Lyngaas Torgersen and Sascha Pust for critically reading the manuscript. This work was supported by grants from the South-Eastern Norway Regional Health Authority, the Norwegian Cancer Society and the Norwegian Research Council through its Centres of Excellence funding scheme, project number 179571.

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Authors and Affiliations

  1. Department of Molecular Cell Biology, Institute for Cancer Research, The Norwegian Radium Hospital, Oslo University Hospital, 0379, Oslo, Norway

    Tove Irene Klokk, Simona Kavaliauskiene & Kirsten Sandvig

  2. Centre for Cancer Biomedicine, Faculty of Medicine, University of Oslo, 0316, Oslo, Norway

    Tove Irene Klokk, Simona Kavaliauskiene & Kirsten Sandvig

  3. Department of Biosciences, University of Oslo, 0316, Oslo, Norway

    Simona Kavaliauskiene & Kirsten Sandvig

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  1. Tove Irene Klokk
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Correspondence to Tove Irene Klokk.

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Klokk, T.I., Kavaliauskiene, S. & Sandvig, K. Cross-linking of glycosphingolipids at the plasma membrane: consequences for intracellular signaling and traffic. Cell. Mol. Life Sci. 73, 1301–1316 (2016). https://doi.org/10.1007/s00018-015-2049-1

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