title: Assays and functional properties of auxilin-dynamin interactions.
abstract: The large GTPase dynamin is required for budding of clathrin-coated vesicles from the plasma membrane, but its mechanism of action is still not understood. Growing evidence indicates that the GTP-bound form of dynamin recruits downstream partners that execute the fission reaction. Recently, we reported nucleotide-dependent interactions between dynamin and auxilin, which suggested that auxilin cooperates with dynamin during vesicle formation. Here we describe three different in vitro assays that monitor auxilin-dynamin interactions, as well as fluorescence lifetime imaging microscopy that identify direct interactions between dynamin and auxilin in cells.
authors.affiliation: Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Sanja
authors.initials: S
authors.lastname: Sever
authors.forename: Jesse
authors.initials: J
authors.lastname: Skoch
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Bacskai
authors.forename: Sherri L
authors.initials: SL
authors.lastname: Newmyer
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D034021:Auxilins / Q000096:biosynthesis / Q000378:metabolism*; D001923:Brain Chemistry; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D022163:Clathrin-Coated Vesicles / Q000502:physiology; D034281:Dynamins / Q000378:metabolism*; D020690:GTPase-Activating Proteins / Q000032:analysis; D005982:Glutathione Transferase / Q000235:genetics; D006801:Humans; D051379:Mice; D008856:Microscopy, Fluorescence; D025941:Protein Interaction Mapping / Q000379:methods*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D018411:Spodoptera
chemical_list: D034021:Auxilins; D020690:GTPase-Activating Proteins; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; D034281:Dynamins