title: Specific interaction of Golgi coatomer protein alpha-COP with phosphatidylinositol 3,4,5-trisphosphate.
abstract: The phosphoinositide binding selectivity of Golgi coatomer COPI polypeptides was examined using photoaffinity analogs of the soluble inositol polyphosphates Ins(1,4,5)P3, Ins(1,3,4,5)P4, and InsP6, and of the polyphosphoinositides PtdIns(3,4,5)P3, PtdIns(4,5)P2, and PtdIns(3,4)P2. Highly selective Ins(1,3,4,5)P4-displaceable photocovalent modification of the alpha-COP subunit was observed with a p-benzoyldihydrocinnamide (BZDC)-containing probe, [3H]BZDC-Ins(1,3,4,5)P4. A more highly phosphorylated probe, [3H]BZDC-InsP6 probe labeled six of the seven subunits, with only beta, beta', delta, and epsilon-COP showing competitive displacement by excess InsP6. Importantly, [3H]BZDC-triester-PtdIns(3,4,5)P3, the lipid with the same phosphorylation pattern as Ins(1,3,4,5)P4, showed specific, PtdIns(3,4,5)P3-displaceable labeling of only alpha-COP. Labeling by the PtdIns(4,5)P2 and PtdIns(3,4)P2 photoaffinity probes was less intense and showed no discrimination based on PtdInsPn ligand. Thus, both the D-3 and D-5 phosphates are critical for the alpha-COP-PtdIns(3,4,5)P3 interaction, suggesting an important role for this polyphosphoinositide in vesicular trafficking.
authors.affiliation: The University of Utah, Department of Medicinal Chemistry, Salt Lake City, Utah 84112-5820, USA.
authors.forename: A
authors.initials: A
authors.lastname: Chaudhary
authors.forename: Q M
authors.initials: QM
authors.lastname: Gu
authors.forename: O
authors.initials: O
authors.lastname: Thum
authors.forename: A A
authors.initials: AA
authors.lastname: Profit
authors.forename: Y
authors.initials: Y
authors.lastname: Qi
authors.forename: L
authors.initials: L
authors.lastname: Jeyakumar
authors.forename: S
authors.initials: S
authors.lastname: Fleischer
authors.forename: G D
authors.initials: GD
authors.lastname: Prestwich
mesh_terms: D000345:Affinity Labels; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D020756:Coatomer Protein; D006056:Golgi Apparatus / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism*
chemical_list: D000345:Affinity Labels; D020756:Coatomer Protein; D008565:Membrane Proteins; D018129:Phosphatidylinositol Phosphates