Calmodulin stimulation and calcium regulation of smooth muscle adenylate cyclase activity

M. T. Piascik, M. Babich, M. E. Rush

Research output: Contribution to journalArticle

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Abstract

The Ca2+-dependent regulation of the adenylate cyclase activity associated with microsomes isolated from bovine aortic smooth muscle has been studied. Calmodulin content of microsomal membranes employed in these studies was 80 ± 14 ng/mg as determined by specific radioimmunoassay. In the absence of exogenous calmodulin, Ca2+ concentrations greater than 0.8 μM inhibited adenylate cyclase activity with one-half-maximal inhibition occurring at 2.5 μM Ca2+. In the presence of 5 or 9 μM bovine testis calmodulin, Ca2+ stimulated smooth muscle adenylate cyclase activity with one-half-maximal stimulation occurring at 0.2 μM for both 5 and 9 μM calmodulin. Calmodulin stimulation was observed between 0.1 and 0.8 μM Ca2+. Despite the presence of calmodulin, Ca2+ concentrations greater than 0.8 μM were inhibitory to smooth muscle adenylate cyclase activity. However, calmodulin reduced the sensitivity of the enzyme to inhibition by Ca2+. Trifluoperazine (100 μM) reversed both the calmodulin-dependent-stimulation of cyclase activity and the calmodulin-induced decrease in sensitivity to the inhibitory actions of Ca2+. Trifluoperazine alone shifted the curve describing Ca2+ inhibition of cyclase activity to the left. The value of Ca2+ for one-half-maximal inhibition decreased from 2.9 to 1.2 μM. The trifluoperazine-induced shift was reversed by exogenous calmodulin. These data suggest: 1) Ca2+, at physiological concentrations, can stimulate as well as inhibit smooth muscle adenylate cyclase activity; 2) the stimulation of adenylate cyclase activity is medidated by calmodulin; 3) the Ca2+-calmodulin complex reduces the sensitivity of smooth muscle adenylate cyclase to the inhibitory actions of Ca2+; and 4) the level of calmodulin associated with smooth muscle adenylate cyclase may modulate the response (both stimulatory and inhibitory) of the enzyme to Ca2+.

Original languageEnglish (US)
Pages (from-to)10913-10918
Number of pages6
JournalJournal of Biological Chemistry
Volume258
Issue number18
StatePublished - 1983
Externally publishedYes

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Calmodulin
Adenylyl Cyclases
Smooth Muscle
Muscle
Calcium
Trifluoperazine
Enzymes
Microsomes
Radioimmunoassay
Testis

ASJC Scopus subject areas

  • Biochemistry

Cite this

Calmodulin stimulation and calcium regulation of smooth muscle adenylate cyclase activity. / Piascik, M. T.; Babich, M.; Rush, M. E.

In: Journal of Biological Chemistry, Vol. 258, No. 18, 1983, p. 10913-10918.

Research output: Contribution to journalArticle

Piascik, MT, Babich, M & Rush, ME 1983, 'Calmodulin stimulation and calcium regulation of smooth muscle adenylate cyclase activity', Journal of Biological Chemistry, vol. 258, no. 18, pp. 10913-10918.
Piascik, M. T. ; Babich, M. ; Rush, M. E. / Calmodulin stimulation and calcium regulation of smooth muscle adenylate cyclase activity. In: Journal of Biological Chemistry. 1983 ; Vol. 258, No. 18. pp. 10913-10918.
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