Characterization of a γ-adaptin ear-binding motif in enthoprotin

• Published in: FEBS letters Vol. 555; no. 3; pp. 437 - 442
• Main Authors: Wasiak, Sylwia, Denisov, Alexei Yu, Han, Zhaozhong, Leventis, Peter A., de Heuvel, Elaine, Boulianne, Gabrielle L., Kay, Brian K., Gehring, Kalle, McPherson, Peter S.
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 18.12.2003
• Subjects: Adaptor Protein Complex gamma Subunits - metabolism; Alanine - genetics; Alanine - metabolism; Amino Acid Motifs - genetics; Amino Acid Sequence; Amino Acid Substitution; Animals; AP-1; AP-2; Binding Sites; Cell Line; Clathrin; Clathrin-coated vesicle; Humans; Mice; Models, Molecular; Molecular Sequence Data; NMR, trans-Golgi network; Nuclear Magnetic Resonance, Biomolecular; Protein Binding; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Sequence Alignment; Transcription Factor AP-1 - chemistry; Transcription Factor AP-1 - metabolism; Transfection; Clathrin; AP-2; AP-1; NMR, trans-Golgi network; Clathrin-coated vesicle
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/S0014-5793(03)01299-7

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 γ-adaptin ear (γ-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 γ-ear in complex with a synthetic peptide from enthoprotin provide structural details of the binding site for TGN accessory proteins within the γ-ear.