Characterization of adenylyl cyclase in goldfish brain.
The rate of production of cAMP by the adenylyl cyclase enzyme from goldfish brain was linear with time and with protein concentration. In agreement with mammalian adenylyl cyclase systems the enzyme is divalent cation dependent, being activated in the presence of either Mg2+ or Mn2+. Forskolin also stimulated the rate of reaction in a dose-dependent manner with a half-maximal effect of 1 microM. The activated enzyme was inhibited by high concentrations of Ca2+ but was independent of Na+ concentration. The presence of guanine nucleotide binding proteins (G-proteins) was demonstrated by the fact that both NaF and guanosine 5'-[beta gamma-imido]triphosphate (p[NH]ppG) stimulated the basal rate. In addition, the p[NH]ppG dose-response curve of the forskolin-stimulated enzyme was biphasic, similar to that observed for other systems. At low concentrations of p[NH]ppG a small inhibition was observed while higher concentrations produced a stimulation. These data suggest that the goldfish brain adenylyl cyclase enzyme complex includes both stimulatory and inhibitory G-proteins in addition to the catalytic unit. A series of known and putative goldfish neurotransmitter substances failed to either stimulate or inhibit the adenylyl cyclase activity. The endogenous neurotransmitters which interact with this second messenger system remain to be determined.
Kirkham
D M
DM
Department of Pharmacology, Medical School, University of Birmingham, Edgbaston.
Henley
J M
JM
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002118:Calcium / Q000494:pharmacology; D002147:Calmodulin / Q000494:pharmacology; D005576:Colforsin; D000242:Cyclic AMP / Q000096:biosynthesis; D006054:Goldfish / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000494:pharmacology; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology
D002147:Calmodulin; D005576:Colforsin; D008345:Manganese; D006160:Guanosine Triphosphate; D000242:Cyclic AMP; D000262:Adenylyl Cyclases; D008274:Magnesium; D002118:Calcium