Internalization of Sambucus nigra agglutinins I and II in insect midgut CF-203 cells

In this project, the uptake mechanisms and localization of two lectins from Sambucus nigra, further referred to as S. nigra agglutinin (SNA)‐I and SNA‐II, into insect midgut CF‐203 cells were studied. SNA‐I is a chimeric lectin belonging to the class of ribosome‐inactivating proteins, whereas SNA‐II is a hololectin devoid of enzymatic activity. Internalization of the fluorescein isothiocyanate‐labeled lectin was investigated using confocal microscopy. Both lectins were internalized into the cytoplasm of CF‐203 cells at similar rates. Preexposure of the insect midgut cells to specific inhibitors of clathrin‐ and caveolae‐mediated endocytosis resulted in an inhibition of lectin uptake in CF‐203 cells and caspase‐induced cytotoxicity caused by SNA‐I and SNA‐II, confirming the involvement of both endocytosis pathways. Further studies demonstrated that the uptake mechanism(s) for both lectins required phosphoinositide 3‐kinases, but did not depend on the actin cytoskeleton. Since the hololectin SNA‐II apparently uses a similar endocytosis pathway as the chimerolectin SNA‐I, it can be concluded that the endocytosis process mainly relies on the carbohydrate‐binding activity of the lectins under investigation. © 2011 Wiley Periodicals, Inc.