Browsing by Subject "clathrin"

Clathrin-mediated endocytosis is the most common pathway by which cells internalize cargoes from the membrane. It is a critical process in cell communication, development, and homeostasis. In order to study endocytic dynamics, it is critical that one can clearly distinguish receptors that have entered the cell from those which remain on the cell membrane. Current techniques for investigating endocytosis rely on removing membrane-bound components with harsh treatments which may interfere with cell physiology, and often depend on antibodies which are not widely available and - even when they are - may give unreliable signals and may affect receptor behavior and internalization rates. Additionally, a large portion of studies on clathrin-mediated endocytosis have been done on a single receptor, the transferrin receptor. Here we have developed a new assay which resolves the above issues through use of a novel protein probe. This fusion protein will allow us to resolve the issues with current endocytic assays mentioned above, and in theory can be used to study any membrane receptor which is endocytosed. Our preliminary results show that we can use our protein to effectively track endocytosed receptors without interference from signal of receptors remaining on the cell membrane. This shows that our protein may be a powerful tool for studying endocytosis across a wide variety of membrane-bound receptors.