title: Cell-free fusion of endocytic vesicles is regulated by phosphorylation.
abstract: Okadaic acid and microcystin-LR, both potent inhibitors of protein phosphatases (PP), blocked vesicle fusion in a cell-free system. The effect of okadaic acid was reversed by the purified catalytic subunit of PP2A, but not PP1. Inhibition was gradual, required Mg-ATP, and was reduced by protein kinase inhibitors, indicating that it was mediated via protein phosphorylation. A candidate protein kinase would be cdc2 kinase, which normally is active in mitotic extracts and has been shown to inhibit endocytic vesicle fusion (Tuomikoski, T., M.-A. Felix, M. Dorée, and J. Gruenberg. 1989. Nature (Lond.). 342:942-945). However, it would appear that cdc2 kinase is not responsible for inhibition by okadaic acid. When compared to cytosol prepared from mitotic cells, okadaic acid did not increase cdc2 kinase activity sufficiently to account for the inhibition. In addition, inhibition was maintained when cdc2 protein was depleted from cytosol.
authors.affiliation: Imperial Cancer Research Fund, Cell Biology Laboratory, London, England.
authors.forename: P G
authors.initials: PG
authors.lastname: Woodman
authors.forename: D I
authors.initials: DI
authors.lastname: Mundy
authors.forename: P
authors.initials: P
authors.lastname: Cohen
authors.forename: G
authors.initials: G
authors.lastname: Warren
mesh_terms: D016203:CDC2 Protein Kinase / Q000378:metabolism; D002474:Cell-Free System; D003600:Cytosol / Q000502:physiology; D004705:Endocytosis; D004988:Ethers, Cyclic / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D066298:In Vitro Techniques; D008387:Marine Toxins; D008561:Membrane Fusion / Q000187:drug effects*; D052998:Microcystins; D019319:Okadaic Acid; D010456:Peptides, Cyclic / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors; D010750:Phosphoproteins / Q000502:physiology*; D014407:Tumor Cells, Cultured
chemical_list: D004988:Ethers, Cyclic; D008387:Marine Toxins; D052998:Microcystins; D010456:Peptides, Cyclic; D010750:Phosphoproteins; D019319:Okadaic Acid; D016203:CDC2 Protein Kinase; D010749:Phosphoprotein Phosphatases; C057862:cyanoginosin LR