title: Analysis of Arf interaction with GGAs in vitro and in vivo.
abstract: Small GTPases of the ADP-ribosylation factor (Arf) family regulate membrane traffic and dynamics in eukaryotic cells. GGAs (Golgi-localizing, gamma-adaptin ear homology domain, Arf-binding proteins) are a family of monomeric clathrin adaptor proteins that were originally identified as proteins interacting with Arfs and found to associate mainly with membranes of the trans-Golgi network (TGN). Like other adaptor and coat proteins, membrane association of GGAs is regulated by Arfs in a GTP-dependent manner. Together with or independent of the adaptor protein complex AP-1, GGAs mediate sorting of transmembrane proteins, including mannose 6-phosphate receptors, between the TGN and endosomes by clathrin-coated vesicles. This chapter describes methods to examine the interaction between Arfs and GGAs and to analyze the cellular function of GGAs regulated by Arfs.
authors.affiliation: Department of Physiological Chemistry, Graduate School of Pharmaceutical Sciences, Kyoto University, Japan.
authors.forename: Kazuhisa
authors.initials: K
authors.lastname: Nakayama
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Takatsu
mesh_terms: D020727:ADP-Ribosylation Factors / Q000378:metabolism*; D033942:Adaptor Proteins, Vesicular Transport / Q000378:metabolism*; D000595:Amino Acid Sequence; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D025941:Protein Interaction Mapping / Q000379:methods; D012441:Saccharomyces cerevisiae; D020798:Two-Hybrid System Techniques
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; C407306:GGA adaptor proteins; D020727:ADP-Ribosylation Factors