title: Interaction of coatomer with aminoglycoside antibiotics: evidence that coatomer has at least two dilysine binding sites.
abstract: Coatomer is the soluble precursor of the COPI coat (coat protein I) involved in traffic among membranes of the endoplasmic reticulum and the Golgi apparatus. We report herein that neomycin precipitates coatomer from cell extracts and from purified coatomer preparations. Precipitation first increased and then decreased as the neomycin concentration increased, analogous to the precipitation of a polyvalent antigen by divalent antibodies. This suggested that neomycin cross-linked coatomer into large aggregates and implies that coatomer has two or more binding sites for neomycin. A variety of other aminoglycoside antibiotics precipitated coatomer, or if they did not precipitate, they interfered with the ability of neomycin to precipitate. Coatomer is know to interact with a motif (KKXX) containing adjacent lysine residues at the carboxyl terminus of the cytoplasmic domains of some membrane proteins resident in the endoplasmic reticulum. All of the antibiotics that interacted with coatomer contain at least two close amino groups, suggesting that the antibiotics might be interacting with the di-lysine binding site of coatomer. Consistent with this idea, di-lysine itself blocked the interaction of antibiotics with coatomer. Moreover, di-lysine and antibiotics each blocked the coating of Golgi membranes by coatomer. These data suggest that certain aminoglycoside antibiotics interact with di-lysine binding sites on coatomer and that coatomer contains at least two of these di-lysine binding sites.
authors.affiliation: Molecular and Cell Biology Program, University of Texas at Dallas, Richardson 75083-0688, USA.
authors.forename: R T
authors.initials: RT
authors.lastname: Hudson
authors.forename: R K
authors.initials: RK
authors.lastname: Draper
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D016466:CHO Cells; D011232:Chemical Precipitation; D020756:Coatomer Protein; D006224:Cricetinae; D004151:Dipeptides / Q000737:chemistry / Q000378:metabolism; D004347:Drug Interactions; D006056:Golgi Apparatus / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism; D008239:Lysine / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D009355:Neomycin / Q000737:chemistry
chemical_list: D000900:Anti-Bacterial Agents; D020756:Coatomer Protein; D004151:Dipeptides; D008565:Membrane Proteins; D009355:Neomycin; D008239:Lysine