The presence of free G protein beta/gamma subunits in human neutrophils results in suppression of adenylate cyclase activity.
We have examined the adenylate cyclase of human neutrophil membranes and compared it to that of human platelet membranes. Stimulated activities were at least 20-fold lower in the neutrophil than in the human platelet. The inhibitory hormone epinephrine was able to attenuate markedly the adenylate cyclase activity of human platelets at micromolar concentrations, whereas little inhibition was observed in the human neutrophil at up to 100 microM concentrations. When we examined the ability of exogenous pure beta/gamma subunits to affect adenylate cyclase activity in both systems, we observed dose-dependent inhibition of stimulated adenylate cyclase activities in the platelet, whereas no inhibition of neutrophil adenylate cyclase could be detected. This difference did not appear to be due to differences in the degree of incorporation of beta/gamma into each membrane. The effects of G protein alpha subunits were also examined. In the platelet, unliganded G protein alpha produced an increase in adenylate cyclase activity of limited extent which saturated at relatively low levels of alpha subunit. In the neutrophil, the effect of unliganded G protein alpha did not appear to saturate and produced much larger relative increases in adenylate cyclase activity. Quantitation of the free beta/gamma activity in neutrophil extracts detected free beta/gamma activity even in the absence of G protein activators. We hypothesize the human neutrophil to be a system in which an excess of free beta/gamma subunits is present and which suppresses neutrophil adenylate cyclase activity. This excess of free beta/gamma minimizes any additional effect of exogenous beta/gamma, but can be reversed by addition of proteins which can bind beta/gamma subunits, e.g. G alpha subunits.
D000067956:Adenylyl Cyclase Inhibitors; D000262:Adenylyl Cyclases / Q000097:blood*; D000818:Animals; D001792:Blood Platelets / Q000201:enzymology; D002417:Cattle; D002462:Cell Membrane / Q000201:enzymology; D004837:Epinephrine / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000097:blood*; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D006801:Humans; D046911:Macromolecular Substances; D009504:Neutrophils / Q000201:enzymology*; D013873:Thionucleotides / Q000494:pharmacology
D000067956:Adenylyl Cyclase Inhibitors; D046911:Macromolecular Substances; D013873:Thionucleotides; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases; D004837:Epinephrine