Abstract
A site-directed photocrosslink approach was used to elucidate components that interact directly with ADP- ribosylation factor (ARF)-GTP during coat assembly. Two ARF mutants were generated that contain a photolabile amino acid at positions distant to each other within the ARF molecule. Here we show that one of the two positions specifically interacts with coatomer subunit beta both on Golgi membranes and in isolated coat protein complex type I (COPI)-coated vesicles. Thus, a direct and GTP-dependent interaction of coatomer via beta-coat protein complex (COP) with ARF is involved in the coating of COPI-coated vesicles. These data implicate a bivalent interaction of the complex with the donor membrane during vesicle formation.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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ADP-Ribosylation Factors
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Affinity Labels
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Azirines
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Biological Transport
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Coated Vesicles / metabolism
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Coatomer Protein
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Endoplasmic Reticulum / metabolism
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GTP-Binding Proteins / genetics
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GTP-Binding Proteins / metabolism*
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GTP-Binding Proteins / radiation effects
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Guanosine 5'-O-(3-Thiotriphosphate) / metabolism
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Guanosine Triphosphate / metabolism*
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Membrane Proteins / metabolism*
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Microtubule-Associated Proteins / metabolism
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Mutation
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Phenylalanine / analogs & derivatives
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Precipitin Tests
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Protein Binding
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Ultraviolet Rays
Substances
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Affinity Labels
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Azirines
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Coatomer Protein
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Membrane Proteins
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Microtubule-Associated Proteins
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Guanosine 5'-O-(3-Thiotriphosphate)
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Phenylalanine
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Guanosine Triphosphate
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3-(4-(3-(trifluoromethyl)-3H-diazirin-3-yl)phenyl)alanine
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GTP-Binding Proteins
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ADP-Ribosylation Factors