Assays and functional properties of auxilin-dynamin interactions

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 repor...

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Bibliographic Details
Published in:	Methods in enzymology Vol. 404; p. 570
Main Authors:	Sever, Sanja, Skoch, Jesse, Bacskai, Brian J, Newmyer, Sherri L
Format:	Journal Article
Language:	English
Published:	United States 2005
Subjects:	Adenoviridae - genetics Animals Auxilins - biosynthesis Auxilins - metabolism Brain Chemistry Cells, Cultured Chromatography, Affinity Clathrin-Coated Vesicles - physiology Dynamins - metabolism Glutathione Transferase - genetics GTPase-Activating Proteins - analysis Humans Mice Microscopy, Fluorescence Protein Interaction Mapping - methods Rats Recombinant Fusion Proteins - biosynthesis Spodoptera
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Summary:	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.
ISSN:	0076-6879
DOI:	10.1016/S0076-6879(05)04050-4