Recruitment of pleckstrin and phosphoinositide 3-kinase gamma into the cell membranes, and their association with G beta gamma after activation of NK cells with chemokines.
The role of phosphoinositide 3 kinases (PI 3-K) in chemokine-induced NK cell chemotaxis was investigated. Pretreatment of NK cells with wortmannin inhibits the in vitro chemotaxis of NK cells induced by lymphotactin, monocyte-chemoattractant protein-1, RANTES, IFN-inducible protein-10, or stromal-derived factor-1 alpha. Introduction of inhibitory Abs to PI 3-K gamma but not to PI 3-K alpha into streptolysin O-permeabilized NK cells also inhibits chemokine-induced NK cell chemotaxis. Biochemical analysis showed that within 2-3 min of activating NK cells, pleckstrin is recruited into NK cell membranes, whereas PI 3-K gamma associates with these membranes 5 min after stimulation with RANTES. Recruited PI 3-K gamma generates phosphatidylinositol 3,4,5 trisphosphate, an activity that is inhibited upon pretreatment of NK cells with wortmannin. Further analysis showed that a ternary complex containing the beta gamma dimer of G protein, pleckstrin, and PI 3-K gamma is formed in NK cell membranes after activation with RANTES. The recruitment of pleckstrin and PI 3-K gamma into NK cell membranes is only partially inhibited by pertussis toxin, suggesting that the majority of these molecules form a complex with pertussis toxin-insensitive G proteins. Our results may have application for the migration of NK cells toward the sites of inflammation.
al-Aoukaty
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A
Department of Anatomy, University of Oslo, Norway.
Rolstad
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B
Maghazachi
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AA
D000730:Androstadienes / Q000494:pharmacology; D001692:Biological Transport / Q000187:drug effects / Q000276:immunology; D001792:Blood Platelets / Q000378:metabolism; D001798:Blood Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D018946:Chemokine CCL5 / Q000494:pharmacology; D018925:Chemokines / Q000494:pharmacology*; D002634:Chemotaxis, Leukocyte / Q000276:immunology; D019281:Dimerization; D004351:Drug Resistance; D019204:GTP-Binding Proteins / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007694:Killer Cells, Natural / Q000187:drug effects / Q000201:enzymology / Q000276:immunology*; D008213:Lymphocyte Activation / Q000276:immunology*; D046911:Macromolecular Substances; D037342:Pertussis Toxin; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism* / Q000502:physiology; D010750:Phosphoproteins; D010566:Virulence Factors, Bordetella / Q000494:pharmacology; D000077191:Wortmannin
D000730:Androstadienes; D001798:Blood Proteins; D018946:Chemokine CCL5; D018925:Chemokines; D046911:Macromolecular Substances; D010750:Phosphoproteins; D010566:Virulence Factors, Bordetella; C039077:platelet protein P47; D037342:Pertussis Toxin; D019869:Phosphatidylinositol 3-Kinases; D019204:GTP-Binding Proteins; D000077191:Wortmannin