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EHDS are serine phosphoproteins: EHD1 phosphorylation is enhanced by serum stimulation


Endocytic processes are mediated by multiple protein-protein interacting modules and regulated by phosphorylation and dephosphorylation. The Eps15 homology domain containing protein 1 (EHD1) has been implicated in regulating recycling of proteins, internalized both in clathrin-dependent and clathrin-independent endocytic pathways, from the recycling compartment to the plasma membrane. EHD1 was found in a complex with clathrin, adaptor protein complex-2 (AP-2) and insulin-like growth factor-1 receptor (IGF-1R), and was shown to interact with Rabenosyn-5, SNAP29, EHBP1 (EH domain binding protein 1) and syndapin I and II. In this study, we show that EHD1, like the other human EHDs, undergoes serine-phosphorylation. Our results also indicate that EHD1 is a serum-inducible serine-phosphoprotein and that PKC (protein kinase C) is one of its kinases. In addition, we show that inhibitors of clathrin-mediated endocytosis decrease EHD1 phosphorylation, while inhibitors of caveolinmediated endocytosis do not affect EHD1 phosphorylation. The results of experiments in which inhibitors of endocytosis were employed strongly suggest that EHD1 phosphorylation occurs between early endosomes and the endocytic recycling compartment.



adaptor protein complex-2


bovine serum albumin


Chinese hamster ovary


counts per minute


dialyzed FCS


Dulbecco modified eagle medium


EH domain binding protein 1


fetal calf serum




insulin-like growth factor-1 receptor


mouse dihydrofolate reductase


minimal essential medium-alpha


open reading frame


phosphate buffered saline


protein kinase C


polyvinidilen difluoride


thin-layer chromatography


thin-layer electrophoresis


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Correspondence to Mia Horowitz.

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Fichtman, B., Ravid, L., Rapaport, D. et al. EHDS are serine phosphoproteins: EHD1 phosphorylation is enhanced by serum stimulation. Cell Mol Biol Lett 13, 632–648 (2008).

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Key words

  • Endocytosis
  • EHD1
  • EH domain
  • Phosphorylation