Characterization of a gamma-adaptin ear-binding motif in enthoprotin |
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| 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 |
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| Affiliation: | Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, 3801 University St., Montreal, QC, Canada H3A 2B4. peter.mcpherson@mcgill.ca |
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| 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. |
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| Keywords: | AP-1 AP-2 Clathrin Clathrin-coated vesicle NMR, trans-Golgi network |
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