We have used 3 main methods to disrupt endocytosis in cells: use of inhibitory peptides, overexpression of protein domains and RNAi. Each method has advantages and disadvantages and thus it is preferable if a number of approaches can be used. We first used the peptide approach in Marks et al 1998 where a dynamin peptide was made membrane permeable and used to inhibit synaptic vesicle recycling. This peptide is now available from Tocris. We are currently developing more potent tools.
In the overexpressions shown below (and elsewhere on the web site), dynamin mutants will inhibit many different endocytic events, while AP180 C-terminus will inhibit clathrin-dependent budding events, while epsin1 R63L+H73L will inhibit AP2 budding events, and epsinR will inhibit AP1 budding events (shown by the inhibition of cathepsinD trafficking to the lysosome in JCB paper).