title: Characterization of a gamma-adaptin ear-binding motif in enthoprotin.
abstract: Enthoprotin, a newly identified component of clathrin-coated vesicles, interacts with the trans-Golgi network (TGN) clathrin adapters AP-1 and GGA2. Here we perform a multi-faceted analysis of the site in enthoprotin that is responsible for the binding to the gamma-adaptin ear (gamma-ear) domain of AP-1. Alanine scan mutagenesis and nuclear magnetic resonance (NMR) studies reveal the full extent of the site as well as critical residues for this interaction. NMR studies of the gamma-ear in complex with a synthetic peptide from enthoprotin provide structural details of the binding site for TGN accessory proteins within the gamma-ear.
authors.affiliation: Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, 3801 University St., Montreal, QC, Canada H3A 2B4. peter.mcpherson@mcgill.ca
authors.forename: Sylwia
authors.initials: S
authors.lastname: Wasiak
authors.forename: Alexei Yu
authors.initials: AY
authors.lastname: Denisov
authors.forename: Zhaozhong
authors.initials: Z
authors.lastname: Han
authors.forename: Peter A
authors.initials: PA
authors.lastname: Leventis
authors.forename: Elaine
authors.initials: E
authors.lastname: de Heuvel
authors.forename: Gabrielle L
authors.initials: GL
authors.lastname: Boulianne
authors.forename: Brian K
authors.initials: BK
authors.lastname: Kay
authors.forename: Kalle
authors.initials: K
authors.lastname: Gehring
authors.forename: Peter S
authors.initials: PS
authors.lastname: McPherson
mesh_terms: D033982:Adaptor Protein Complex gamma Subunits / Q000378:metabolism*; D000409:Alanine / Q000235:genetics / Q000378:metabolism; D020816:Amino Acid Motifs / Q000235:genetics*; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D018808:Transcription Factor AP-1 / Q000737:chemistry / Q000378:metabolism; D014162:Transfection
chemical_list: D033982:Adaptor Protein Complex gamma Subunits; D011994:Recombinant Proteins; D018808:Transcription Factor AP-1; D000409:Alanine