Ca2+-dependent regulation of guinea pig brain adenylate cyclase

M. T. Piascik, P. L. Wisler, C. L. Johnson, J. D. Potter

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91 Scopus citations


The effect of Ca2+ on guinea pig brain adenylate cyclase has been studied using metal buffers to regulate the free Ca2+ concentration. Particulate fractions prepared in the presence of Ca2+ (to bind all the available calmodulin to all of the available binding sites) exhibited a biphasic response to Ca2+. The first was a 2-fold stimulation of basal cyclase activity (half-maximal =0.08 μm). At higher [Ca2+], the cyclase was inhibited to -80 to 90% of basal (half-maximal inhibition=0.3 μM). Particulate fractions prepared by exhaustive washing with EDTA and ethylene glycol bis(β-aminoethyl ether)N,N,N',N'-tetraacetic acid (EGTA) (in an attempt to deplete the membranes of calmodulin) exhibited no stimulation of adenylate cyclase by Ca2+ but did show the Ca2+-dependent inhibitory phase (halfmaximal inhibition=0.5 μM). The calmodulin content of this preparation (EGTA-membranes) was 0.24 μg/mg of protein. The activation phase could be restored to EGTA-membranes by addition of calmodulin. The Ca2+ dependence of calmodulin-dependent phosphodiesterase exhibited a half-maximal activation at [Ca2+]=0.3 μM, similar to the half-maximal inhibition of cyclase in the two preparations. Likewise, Sr2+ activation of phosphodiesterase (half-maximal stimulation=0.1 mM) occurred over the same [Sr2+] range as did half-maximal inhibition of cyclase (0.3 mM) in EGTA-and Ca-membranes. The Ca2+ and Sr2+ dependence of calmodulin tyrosine fluorescence (an indirect measure of Ca2+ and Sr2+ dependence of phosphodiesterase activation and the inhibition of the cyclase. These results suggest that the Ca2+ dependent inhibition of adenylate cyclase may be mediated either by calmodulin or by a metal binding site which has binding properties similar to calmodulin. Thus, our results show that there is an inverse relationship between adenylate cyclase activity and phosphodiesterase activity and suggest that intracellular calcium may indicate a coordinated regulation of the rates of synthesis and degradation of cyclic nucleotides.

Original languageEnglish
Pages (from-to)4176-4181
Number of pages6
JournalJournal of Biological Chemistry
Issue number9
StatePublished - 1980

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology


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