Hydrolysis-resistant GTP analogs stimulate catecholamine release from digitonin-permeabilized PC12 cells.
The effect of the hydrolysis-resistant GTP analogs, guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) and guanylyl imidodiphosphate (GMPPNP), on norepinephrine (NE) secretion from digitonin-permeabilized rat pheochromocytoma cells, PC12, was examined. Although secretion in the presence of saturating Ca2+ (10 microM) was not affected by GTP gamma S or GMPPNP, secretion in the absence of Ca2+ was stimulated by these GTP analogs. Secretion induced by saturating concentrations of GTP gamma S or GMPPNP was approximately 80% of that induced by 10 microM Ca2+. Half-maximum stimulation was induced by 30 microM GTP gamma S or GMPPNP. Both Ca2(+)-stimulated and GTP gamma S-stimulated secretion were ATP dependent and inhibited by N-ethylmaleimide. The GTP gamma S-stimulated secretion of NE from permeabilized PC12 cells does not appear to result from either the release of Ca2+ or the activation of protein kinase C. Activation of protein kinase C by pretreatment of intact cells with 12-O-tetradecanoylphorbol 13-acetate caused a 50% increase in both Ca2(+)-stimulated and GTP gamma S-stimulated secretion. Cholera and pertussis toxins did not affect Ca2(+)-stimulated or GTP gamma S-stimulated NE secretion. Guanosine 5'-O-(2-thiodiphosphate) (GDP beta S) and GTP inhibited GTP gamma S-stimulated secretion but not Ca2(+)-stimulated secretion. The inability of GDP beta S to inhibit Ca2(+)-stimulated secretion indicates that the process affected by GTP gamma S is not an essential step in the Ca2(+)-stimulated pathway.
Carroll
A G
AG
Lombardi Cancer Research Center, Georgetown University Medical Center, Washington, DC.
Rhoads
A R
AR
Wagner
P D
PD
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000310:Adrenal Gland Neoplasms / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002463:Cell Membrane Permeability / Q000187:drug effects; D004072:Digitonin / Q000494:pharmacology*; D005033:Ethylmaleimide / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000037:antagonists & inhibitors / Q000502:physiology; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate / Q000031:analogs & derivatives* / Q000494:pharmacology; D006165:Guanylyl Imidodiphosphate / Q000494:pharmacology*; D006868:Hydrolysis; D007700:Kinetics; D009638:Norepinephrine / Q000378:metabolism*; D010673:Pheochromocytoma / Q000378:metabolism*; D051381:Rats; D015398:Signal Transduction; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013873:Thionucleotides / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D013873:Thionucleotides; D006165:Guanylyl Imidodiphosphate; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D019204:GTP-Binding Proteins; D004072:Digitonin; D013755:Tetradecanoylphorbol Acetate; D005033:Ethylmaleimide; D002118:Calcium; D009638:Norepinephrine