Localization of cyclic nucleotide phosphodiesterase 2 in the brain-derived Triton-insoluble low-density fraction (raft).
The cyclic nucleotides perform a variety of roles in the formation and remodeling of the neuronal interaction. The membrane microdomain called "raft" has been paid much attention, for this domain contains many signal-transducing molecules including trimeric G proteins and cytoskeletal proteins. The raft domain is recovered in a low-density fraction after the treatment of the membrane with a non-ionic detergent such as Triton X-100. The enrichment of cholesterol and sphingolipids is ascribed to be responsible for the detergent insolubility. In this study we focused on the cyclic nucleotide signaling process in rafts prepared from the cerebral cortex of 10-day-old rat and the synaptic plasma membrane fraction and found the presence of a high cAMP and cGMP phosphodiesterase (PDE) activity. The activity was effectively inhibited with erythro-9-(2-hydroxy-3-nonyl)adenine, a PDE2-specific inhibitor but not with other inhibitors such as vinpocetine, quazione, or zaprinast. Further western blotting analysis confirmed the localization of PDE2 in the raft fraction. The presence of adenylyl cyclase V/VI and PKA in the raft fraction was also shown with Western blotting. These results suggest the participation of the raft in the cyclic nucleotide signaling cascade in neurons.
Noyama
Kenjiro
K
Division of Bioinformation, Department of Life Science, Graduate School of Science and Technology, Kobe University, Rokkodai 1-1, Nada-ku, Kobe 657-8501, Japan.
Maekawa
Shohei
S
D000818:Animals; D002021:Buffers; D002540:Cerebral Cortex / Q000187:drug effects / Q000201:enzymology*; D054678:Cyclic Nucleotide Phosphodiesterases, Type 2; D004791:Enzyme Inhibitors / Q000494:pharmacology; D021962:Membrane Microdomains / Q000378:metabolism*; D009712:Nucleotides, Cyclic / Q000378:metabolism; D017830:Octoxynol; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D012995:Solubility; D013347:Subcellular Fractions / Q000201:enzymology
D002021:Buffers; D004791:Enzyme Inhibitors; D009712:Nucleotides, Cyclic; D017830:Octoxynol; D010727:Phosphoric Diester Hydrolases; D054678:Cyclic Nucleotide Phosphodiesterases, Type 2; C517278:Pde2a protein, rat