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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adaptor Protein Complex gamma Subunits / metabolism*
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Alanine / genetics
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Alanine / metabolism
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Amino Acid Motifs / genetics*
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Amino Acid Sequence
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Amino Acid Substitution
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Animals
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Binding Sites
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Cell Line
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Humans
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Mice
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Models, Molecular
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Molecular Sequence Data
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Nuclear Magnetic Resonance, Biomolecular
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Protein Binding
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Sequence Alignment
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Transcription Factor AP-1 / chemistry
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Transcription Factor AP-1 / metabolism
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Transfection
Substances
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Adaptor Protein Complex gamma Subunits
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Recombinant Proteins
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Transcription Factor AP-1
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Alanine