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PMID: 1309811 Published · ppublish English Journal Article

Cell-free fusion of endocytic vesicles is regulated by phosphorylation.

The Journal of cell biology ·Vol. 116 ·No. 2 ·1992-01-00 ·Pages 331-8

Woodman PG, Mundy DI, Cohen P, Warren G

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.

MeSH Terms
CDC2 Protein Kinase/metabolism Cell-Free System Cytosol/physiology Endocytosis Ethers, Cyclic/pharmacology HeLa Cells Humans In Vitro Techniques Marine Toxins Membrane Fusion/drug effects Microcystins Okadaic Acid Peptides, Cyclic/pharmacology Phosphoprotein Phosphatases/antagonists & inhibitors Phosphoproteins/physiology Tumor Cells, Cultured
Chemicals
Ethers, Cyclic Marine Toxins Microcystins Peptides, Cyclic Phosphoproteins Okadaic Acid CDC2 Protein Kinase Phosphoprotein Phosphatases cyanoginosin LR
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Woodman P G
Imperial Cancer Research Fund, Cell Biology Laboratory, London, England.
Mundy D I
Cohen P
Warren G
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.116.2.331
Published
1992-01-00
Pages
331-8
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2289299
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.116.2.331 PMC Full Text View journal details
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