Hormonally stimulated adenylate cyclase and cAMP dependent protein kinase in membranes of rabbit erythroid cells separated according to density.
Plasma membranes were prepared after density gradient separation of erythroid cells obtained from bled animals. In a fraction enriched in young reticulocytes (lowest density), the basal and the prostaglandin stimulated adenylate cyclase were greatly augmented if compared with the membranes from unfractionated cells or from the layers of higher densities. Potentiation by GTP or soluble factor(s) of the prostaglandin stimulated adenylate cyclase was found solely in the fractions containing the youngest cells (lowest density). A very significant augmentation of both the basal and the effectors stimulated adenylate cyclase was obtained when the white blood cells and the platelets were removed by filtration through alpha-cellulose prior to density separation. A small population of probably very young reticulocytes was shown to contain a very active adenylate cyclase coupled to the hormonal receptor. Upon short time of maturation this coupling could no longer be detected. The cAMP generated in the fraction enriched in young reticulocytes increased the phosphorylation of some membrane proteins. The presence of a hormonally regulated adenylate cyclase and eventually the phosphorylation of some specific membrane proteins by the cAMP generated in situ permit to envisage possible functions of this system in young reticulocytes.
D000262:Adenylyl Cyclases / Q000097:blood*; D000527:Alprostadil; D000818:Animals; D002499:Centrifugation, Density Gradient; D000242:Cyclic AMP / Q000494:pharmacology; D004789:Enzyme Activation; D004910:Erythrocyte Membrane / Q000201:enzymology*; D004912:Erythrocytes / Q000201:enzymology*; D006160:Guanosine Triphosphate / Q000494:pharmacology; D011458:Prostaglandins E / Q000494:pharmacology*; D011494:Protein Kinases / Q000097:blood*; D011817:Rabbits; D012156:Reticulocytes / Q000201:enzymology
D011458:Prostaglandins E; D006160:Guanosine Triphosphate; D000242:Cyclic AMP; D011494:Protein Kinases; D000262:Adenylyl Cyclases; D000527:Alprostadil