3] Detection of endocytic vesicle fusion in Vitro, using assay based on avidin-biotin association reaction

Abstract

Publisher Summary Endocytosis and phagocytosis are the two primary mechanisms employed by eukaryotic cells to internalize macromolecules from the extracellular milieu. The most comprehensively studied endocytic pathway is receptor-mediated endocytosis via clathrin-coated pits and subsequent transport to lysosomes. In the early endosome, ligandreceptor dissociation and sorting occur. Soon afterward, receptors recycle to the plasma membrane, while free ligand progresses to another population of vesicles termed, “late endosomes.” Contrary to endocytosis, phagocytosis is a function of specialized cells. In vertebrates, mononuclear macrophages and polymorphonuclear granulocytes are the primary phagocytes. The biochemical mechanisms that regulate phagolysosome biogenesis are very complex. As evidenced by a number of investigations, both membrane fusion and membrane recycling play important roles in this process. Therefore, to better understand the mechanisms employed by phagosomes to process and traffic internalized material, development of probes and cell-free assays.