title: [Coupling of GTP-binding protein to inositol phospholipid metabolism in chemoattractant-stimulated guinea pig peritoneal exudate macrophages].
abstract: The involvement of GTP-binding protein in inositol phospholipid metabolism in guinea pig peritoneal exudate macrophages stimulated with the chemoattractant N-formyl-Methionyl-Leucyl-Phenylalanine (fMLP) was examined. The GTP analog, guanosine 5'-[gamma-thio]triphosphate (GTP gamma S) caused a dose-dependent increase in the formation of inositol triphosphate in membranes of macrophages. This effect was specific for GTP and its analog. fMLP-induced inositol phospholipid turnover was markedly inhibited by the prior exposure of macrophages to 100 ng/ml of pertussis toxin (PT). Likewise, the pretreatment of macrophages with 100 ng/ml of PT evoked the inhibition of the increase in the intracellular free Ca2+ concentration and the spreading of macrophages induced by fMLP. These actions of PT were not associated with an alteration in the cellular concentration of cyclic AMP. Incubation of the membranes of macrophages with [32P]NAD and PT resulted in the ADP-ribosylation of a 41,000 Da protein. This ADP-ribosylation was diminished by the prior incubation of the membranes with 100 microM GTP gamma S plus 1 mM MgCl2, indicating that the 41,000 Da protein may be the alpha subunit of a GTP-binding protein. Moreover, there was a parallel between the time course of the ADP-ribosylation of intact macrophages by PT and the inhibition of the increase in intracellular free Ca2+ concentration as well as of the enhancement of the spreading of macrophages. These results suggest that the 41,000 Da protein, a GTP-binding protein, mediates the fMLP-stimulated inositol phospholipid metabolism.
authors.forename: M
authors.initials: M
authors.lastname: Kukita
mesh_terms: D000818:Animals; D001202:Ascitic Fluid / Q000166:cytology; D019204:GTP-Binding Proteins / Q000502:physiology*; D006168:Guinea Pigs; D008264:Macrophages / Q000378:metabolism*; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000378:metabolism; D010716:Phosphatidylinositols / Q000096:biosynthesis*
chemical_list: D010716:Phosphatidylinositols; D009240:N-Formylmethionine Leucyl-Phenylalanine; D019204:GTP-Binding Proteins