Cyclic Nucleotides Cyclic Nucleotide Phosphodiesterase Protein Kinase And Phosphoprotein Phosphatase In The Central Nervous System Of Manduca Sexta

Adenosine 3', 5' -cyclic monophosphate (cyclic AMP, cAMP) and guanosine 3', 5' -cyclic monophosphate (cyclic GMP, cGMP) were quantitated in the central nervous system (CNS) of the insect Manduca sexta by competitive protein binding techniques. The M. sexta CNS was found to contain a strikingly high level of cGMP, about 100-fold greater than that of mammalian brain. The level of cAMP, however, was estimated to be about one-sixth that of mammalian brain. The basal ratio of cGMP/cAMP in the insect CNS was approximately ten, whereas this ratio has been reported to be less than one in a variety of vertebrate and invertebrate tissues. Acetylcholine caused a great elevation of cGMP, but not cAMP, in the M. sexta CNS. Short-term incubation with ecdysterone (insect metamorphosis hormone) promoted the accumulation of both cyclic nucleotides. The existence of cyclic nucleotide- stimulable protein kinases (ATP: protein phosphotransferase, EC 2. 7. 1. 37), a system of enzymes postulated to be instrumental in the biochemical expression of cAMP and cGMP, was demonstrated in the CNS of both larval and adult M. sexta. At low concentrations, cAMP was a much more effective activator of kinase activity than cGMP, Cyclic AMP lowered the Km of the CNS kinase for ATP, a phenomenon which has been reported to be unique to nervous tissue in mammals. A number of the enzymological properties of the insect kinase were similar to those reported in the literature for this enzyme in mammalian tissues. This insect CNS was also shown to possess a potent enzyme system, viz. phosphoprotein phosphatase, for the dephosphorylation of the phosphorylated products of kinase. Phosphatase activity was gauged using phosphoprotamine as substrate. Both kinase and phosphatase activities were found to be enriched in particulate fractions of the CNS. An enzyme system for the destruction of cAMP and cGMP, cyclic nucleotide phosphodiesterase (nucleoside 3', 5' -cyclic phosphate nucleoside 5' -phosphate 31 -phosphohydrolase, EC 3.1. 4