Somatostatin induces release of the alpha subunits of pertussis toxin-sensitive G proteins in native membranes and in intact GH4C1 rat pituitary cells.
Incubation of GH4C1 rat pituitary cell membranes with the poorly hydrolyzable GTP analogue, GTP gamma S, produces a decrease in the pertussis toxin-catalyzed ADP-ribosylation of 40-kDa protein in the membrane pellet and the release of an alpha-like substrate from the membrane into the supernatant fraction; these effects do not occur with the inactive GDP analogue, GDP beta S. The resolved supernatant fraction from GTP gamma S-stimulated membranes is significantly activated to pertussis toxin-catalyzed [32P]ADP-ribosylation by the addition of purified beta gamma complex. Immunoblot analysis identifies the released pertussis toxin substrate as alpha subunits of Gi2, Gi3, and G(o) in the resolved supernatant. The physiological agonist, somatostatin, also stimulates the release of Gi2 and G(o) alpha subunits but not Gi3 from GH4C1 cell membranes in the presence of a low concentration of GTP gamma S (20 nM). The effects of somatostatin are inhibited by pretreatment of GH4C1 cells with pertussis toxin. Furthermore, the addition of somatostatin to intact GH4C1 cells decreases the level of Gi2 alpha subunits in the crude membrane whereas immunoblot analysis of the 274,000 x g supernatant (cytosolic fraction) clearly shows the presence of Gi2 alpha subunits. These data indicate that pertussis toxin-sensitive G proteins in GH4C1 cells dissociate into alpha subunits and beta gamma complex with the release of the alpha subunits from the membranes upon somatostatin activation.
Yajima
Y
Y
Department of Molecular Biology, Tokyo Metropolitan Institute of Medical Science, Japan.
Akita
Y
Y
Katada
T
T
Saito
T
T
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D000818:Animals; D002462:Cell Membrane / Q000187:drug effects; D019204:GTP-Binding Proteins / Q000187:drug effects*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D006153:Guanosine Diphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D037342:Pertussis Toxin; D010903:Pituitary Gland, Anterior / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D017481:Receptors, Somatostatin / Q000187:drug effects* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013004:Somatostatin / Q000494:pharmacology*; D013873:Thionucleotides / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D017481:Receptors, Somatostatin; D013873:Thionucleotides; D010566:Virulence Factors, Bordetella; D006153:Guanosine Diphosphate; D000246:Adenosine Diphosphate Ribose; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D013004:Somatostatin; C023427:guanosine 5'-O-(2-thiodiphosphate); D037342:Pertussis Toxin; D019204:GTP-Binding Proteins