EpsinR D34G + R67L overexpression in COS cells
This pair of mutations abolishes PtdInsP binding
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Myc-EpsinR D34G + R67L with endogenous AP1 and internalised transferrin in COS cells
This mutant has a different distribution to WT epsinR, being found in more peripheral puncta. This shows that the ENTH domain is essential for efficient TGN targeting. The co-localisation images (see below) do not identify the labelled compartment, but there is a redistribution of perinuclear clathrin into these puncta and also a redistribution of TGN46 into these puncta. As seen on the left, AP1 and internalised transferrin do not co-localise. Click image to view individual channels or proceed to galleries of images below. |
Colocalisation of EpsinR D34G + R67L (green) with markers (red), focusing on the perinuclear region.
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Clathrin |
M6P receptor |
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GM130 |
TGN46 |
Blue - Transferrin |
Comparison of epsin1 and epsinR
Myc-epsinR (WT) overexpression and colocalisation with markers
Myc-epsinR D34G+R67L (lipid binding mutant)
Myc-epsinR L10E (proposed lipid conformation mutant)
Myc-epsinR D342R+D422R (clathrin binding mutant)
Myc-epsinR D34G+R67L + D342R+D422R (lipid and clathrin binding mutant)