Cytosolic phospholipase A2 (cPLA2)–generated arachidonic acid (AA) has been shown to be an essential requirement for the activation of NADPH oxidase, in addition to its being the major enzyme involved in the formation of eicosanoid at the nuclear membranes. The mechanism by which cPLA2 regulates NADPH oxidase activity is not known, particularly since the NADPH oxidase complex is localized in the plasma membranes of stimulated cells. The present study is the first to demonstrate that upon stimulation cPLA2 is transiently recruited to the plasma membranes by a functional NADPH oxidase in neutrophils and in granulocyte-like PLB-985 cells. Coimmunoprecipitation experiments and double labeling immunofluorescence analysis demonstrated the unique colocalization of cPLA2 and the NADPH oxidase in plasma membranes of stimulated cells, in correlation with the kinetic burst of superoxide production. A specific affinity in vitro binding was detected between GST-p47phox or GST-p67phox and cPLA2 in lysates of stimulated cells. The association between these two enzymes provides the molecular basis for AA released by cPLA2 to activate the assembled NADPH oxidase. The ability of cPLA2 to regulate two different functions in the same cells (superoxide generation and eicosanoid production) is achieved by a novel dual subcellular localization of cPLA2 to different targets.
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18 August 2003
Article|
August 11 2003
Unique targeting of cytosolic phospholipase A2 to plasma membranes mediated by the NADPH oxidase in phagocytes
Zeev Shmelzer,
Zeev Shmelzer
1Infectious Diseases Laboratory, Department of Clinical Biochemistry
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Nurit Haddad,
Nurit Haddad
1Infectious Diseases Laboratory, Department of Clinical Biochemistry
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Ester Admon,
Ester Admon
1Infectious Diseases Laboratory, Department of Clinical Biochemistry
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Itai Pessach,
Itai Pessach
1Infectious Diseases Laboratory, Department of Clinical Biochemistry
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Thomas L. Leto,
Thomas L. Leto
3Laboratory of Host Defenses, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892
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Zahit Eitan-Hazan,
Zahit Eitan-Hazan
1Infectious Diseases Laboratory, Department of Clinical Biochemistry
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Michal Hershfinkel,
Michal Hershfinkel
2Department of Physiology, Faculty of Health Sciences, Soroka Medical Center and Ben-Gurion University of the Negev, Beer Sheva 84105, Israel
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Rachel Levy
Rachel Levy
1Infectious Diseases Laboratory, Department of Clinical Biochemistry
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Zeev Shmelzer
1Infectious Diseases Laboratory, Department of Clinical Biochemistry
Nurit Haddad
1Infectious Diseases Laboratory, Department of Clinical Biochemistry
Ester Admon
1Infectious Diseases Laboratory, Department of Clinical Biochemistry
Itai Pessach
1Infectious Diseases Laboratory, Department of Clinical Biochemistry
Thomas L. Leto
3Laboratory of Host Defenses, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892
Zahit Eitan-Hazan
1Infectious Diseases Laboratory, Department of Clinical Biochemistry
Michal Hershfinkel
2Department of Physiology, Faculty of Health Sciences, Soroka Medical Center and Ben-Gurion University of the Negev, Beer Sheva 84105, Israel
Rachel Levy
1Infectious Diseases Laboratory, Department of Clinical Biochemistry
Address correspondence to Rachel Levy, Infectious Diseases Laboratory, Dept. of Clinical Biochemistry, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva 84105, Israel. Tel.: 972-8-6403186. Fax: 972-8-6467477. email: [email protected]
Abbreviations used in this paper: AA, arachidonic acid; cPLA2, cytosolic phospholipase A2; fMLP, formyl-methionyl-leucyl-phenylalanine; OZ, opsonized zymosan; PtdIns(4,5)P2, phosphatidylinositol 4,5-bisphosphate.
Received:
November 15 2002
Accepted:
May 09 2003
Online ISSN: 1540-8140
Print ISSN: 0021-9525
The Rockefeller University Press
2003
J Cell Biol (2003) 162 (4): 683–692.
Article history
Received:
November 15 2002
Accepted:
May 09 2003
Citation
Zeev Shmelzer, Nurit Haddad, Ester Admon, Itai Pessach, Thomas L. Leto, Zahit Eitan-Hazan, Michal Hershfinkel, Rachel Levy; Unique targeting of cytosolic phospholipase A2 to plasma membranes mediated by the NADPH oxidase in phagocytes . J Cell Biol 18 August 2003; 162 (4): 683–692. doi: https://doi.org/10.1083/jcb.200211056
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