Inhibition of endocytosis by anti-clathrin antibodies

We examined the function of clathrin, a cytoplasmic protein associated with coated pits and vesicles, by introducing monoclonal antibodies into living cells and determining their effects on membrane transport. When anti-clathrin heavy chain antibodies were used, the following effects were observed: clathrin became aggregated in the cytoplasm, the number of coated pits on the plasma membrane was reduced, and adsorbtive endocytosis of Semliki Forest virus and fluid-phase endocytosis were decreased by 40%-50%. No change in transport of newly synthesized influenza hemagglutinin to the plasma membrane was observed. The results indicated that clathrin in CV-1 cells is involved in fluid-phase uptake and receptor-mediated endocytosis, but not in constitutive transport within the secretory pathway.