GTP-binding proteins associated with serotonin-activated adenylate cyclase in Fasciola hepatica.
The liver fluke Fasciola hepatica has serotonin (5-hydroxytryptamine) receptors that function through a transmembrane signalling system requiring GTP which activates adenylate cyclase (ATP pyrophosphate-lyase (cyclising), EC 4.6.1.1). Non-hydrolysable GTP analogs and NaF activate adenylate cyclase in membrane particles of these organisms. The nature of GTP-binding proteins in these membranes was studied using bacterial toxins and photoaffinity labelling. Treatment of membrane particles from flukes with cholera toxin increased basal adenylate cyclase activity, but markedly decreased activation by serotonin, non-hydrolysable GTP analogs, and NaF. [32P]ADP-ribosylation by cholera toxin or photoaffinity labelling with [32P]-8-N3GTP identified a 53 kDa protein and a 45 kDa protein which appeared to be similar to the forms of the alpha-subunit of the GTP-binding protein associated with adenylate cyclase in mammals. Treatment of membrane particles by pertussis toxin did not significantly change basal adenylate cyclase activity and did not change the stimulation of cyclase by activators. A 43 kDa protein which was [32P]ADP-ribosylated by either cholera or pertussis toxin, depending on the conditions used, and photoaffinity labelled by [32P]-8-N3GTP may be part of the transmembrane signalling system in the liver flukes.
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D037361:Adenylate Cyclase Toxin; D000262:Adenylyl Cyclases / Q000378:metabolism*; D000345:Affinity Labels; D000818:Animals; D001345:Autoradiography; D002772:Cholera Toxin; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D005210:Fasciola hepatica / Q000032:analysis* / Q000201:enzymology / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000032:analysis* / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000494:pharmacology; D037342:Pertussis Toxin; D012701:Serotonin / Q000378:metabolism / Q000494:pharmacology*; D010566:Virulence Factors, Bordetella
D037361:Adenylate Cyclase Toxin; D000345:Affinity Labels; D010566:Virulence Factors, Bordetella; D000246:Adenosine Diphosphate Ribose; D012701:Serotonin; D006160:Guanosine Triphosphate; D002772:Cholera Toxin; D037342:Pertussis Toxin; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases