Claudin-derived peptides are internalized via specific endocytosis pathways

Claudin proteins are involved in the paracellular tightening of epithelia and endothelia. Their internalization, which can be modulated by extracellular stimuli, for example, proinflammatory cytokines, is a prerequisite for the regulation of the paracellular barrier to allow, for instance, cell migration or drug delivery. The internalization of peptide sequences of claudins is completely unknown. Here, we studied the internalization of two peptides, TAMRA-claudin-1 and TAMRA-claudin-5, derivatives of the extracellular loop of claudin-1 and -5, respectively, in either epithelial or endothelial cells. The cellular uptake of the claudin-1 peptide follows the clathrin-mediated endocytosis as indicated by inhibitors and respective tracers for colocalization. In addition, macropinocytosis and caveolae-mediated endocytosis of the peptide was observed. In contrast, the claudin-5 peptide is mainly internalized via the caveolae-mediated endocytosis evidenced by the colocalization with respective tracers and vesicle markers, whereas the nonselective macropinocytosis seems to be involved in a less effective manner. In conclusion, the assumption is supported that claudin peptides can be internalized by specific and nonspecific pathways.