Guanine nucleotide binding proteins mediate the chemotactic signal of transforming growth factor-beta 1 in rat IL-2 activated natural killer cells.
Transforming growth factor (TGF)-beta 1 induced rat IL-2-activated natural killer (IANK) cell chemotaxis. Various doses of cholera toxin (CT) or pertussis toxin (PT) inhibited the activity of TGF-beta 1 suggesting a role for guanine nucleotide binding (G) proteins. ADP-ribosylation assay showed that rat IANK cell membranes possess a 39 kDa PT substrate and two, 41 and 42 kDa, CT substrates. ADP-ribosylation also showed that incubating IANK cell membranes with TGF-beta 1 in the presence of guanosine 5'-O-(3-thiotriphosphate) resulted in the disappearance of the PT substrate. Immunoblot analysis showed that rat IANK cell membranes possess one Gi (39 kDa), one G0 (39 kDa) and three Gs (40, 41, and 42 kDa) proteins. Pretreatment of IANK cell membranes with TGF-beta 1 in the presence of guanosine-5'-O-(3-thiotriphosphate) reduced the intensity of the 39 kDa G(0) and the 40 kDa Gs but not the 39 kDa Gi or the 41 kDa or 42 kDa Gs. Furthermore, TGF-beta 1 stimulated GTP binding and increased GTPase activity in IANK cell membranes. Both activities were inhibited by PT or CT. This inhibition was associated with the modification of G proteins by the toxins suggesting that bacterial toxin substrates are linked to TGF-beta 1 receptors. Our results suggest that G0 and Gs are involved in mediating the chemotactic signal of TGF-beta 1 in rat IANK cells.
Northeastern Ontario Regional Cancer Centre, Sudbury, Canada.
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Maghazachi
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al-Aoukaty
D000818:Animals; D001427:Bacterial Toxins / Q000494:pharmacology; D002478:Cells, Cultured; D002634:Chemotaxis, Leukocyte; D020558:GTP Phosphohydrolases / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000502:physiology*; D006160:Guanosine Triphosphate / Q000378:metabolism; D007376:Interleukin-2 / Q000494:pharmacology*; D007694:Killer Cells, Natural / Q000276:immunology*; D008213:Lymphocyte Activation; D008297:Male; D051381:Rats; D011916:Rats, Inbred F344; D018125:Receptors, Transforming Growth Factor beta / Q000032:analysis; D013047:Specific Pathogen-Free Organisms; D016212:Transforming Growth Factor beta / Q000494:pharmacology*
D001427:Bacterial Toxins; D007376:Interleukin-2; D018125:Receptors, Transforming Growth Factor beta; D016212:Transforming Growth Factor beta; D006160:Guanosine Triphosphate; D020558:GTP Phosphohydrolases; D019204:GTP-Binding Proteins