Oxidation of critical cysteine residues of type I adenylyl cyclase by o-iodosobenzoate or nitric oxide reversibly inhibits stimulation by calcium and calmodulin

Roy J. Duhe, Mark D. Nielsen, Andrew H. Dittman, Enrique C. Villacres, Eui Ju Choi, Daniel R. Storm

Research output: Contribution to journalArticle

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Abstract

The calmodulin binding domain of the type I adenylyl cyclase has recently been identified as an amino acid sequence (residues 495-522) that contains 2 cysteine residues. Therefore, we examined the effect of several sulfhydryl reagents on the calmodulin sensitivity of the enzyme. Treatment of membranes containing the type I adenylyl cyclase with N-ethylmaleimide rapidly inhibited basal, calcium/calmodulin-stimulated, and for-skolin-stimulated adenylyl cyclase activity. When the enzyme was treated with limiting amounts of o-iodosobenzoate, which oxidizes vicinal sulfhydryls to disulfides, stimulation by Ca2+ and calmodulin was eliminated at concentrations which did not affect basal adenylyl cyclase activity. Calmodulin stimulation of the enzyme was restored by treatment with dithiothreitol or glutathione which reduce disulfides to free thiols. NO and sodium nitroprusside also reversible inhibited calmodulin stimulation of the enzyme. We propose that the loss in calmodulin sensitivity caused by these reagents may be due to the oxidation one or more sets of vicinal thiols present in the enzyme.

Original languageEnglish
Pages (from-to)7290-7296
Number of pages7
JournalJournal of Biological Chemistry
Volume269
Issue number10
Publication statusPublished - 1994 Mar 11
Externally publishedYes

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Calmodulin
Cysteine
Nitric Oxide
Calcium
Oxidation
Enzymes
Adenylyl Cyclases
Sulfhydryl Compounds
Disulfides
Sulfhydryl Reagents
Ethylmaleimide
Dithiothreitol
Nitroprusside
adenylyl cyclase 1
Glutathione
Amino Acid Sequence
Membranes
Amino Acids

ASJC Scopus subject areas

  • Biochemistry

Cite this

Oxidation of critical cysteine residues of type I adenylyl cyclase by o-iodosobenzoate or nitric oxide reversibly inhibits stimulation by calcium and calmodulin. / Duhe, Roy J.; Nielsen, Mark D.; Dittman, Andrew H.; Villacres, Enrique C.; Choi, Eui Ju; Storm, Daniel R.

In: Journal of Biological Chemistry, Vol. 269, No. 10, 11.03.1994, p. 7290-7296.

Research output: Contribution to journalArticle

Duhe, Roy J. ; Nielsen, Mark D. ; Dittman, Andrew H. ; Villacres, Enrique C. ; Choi, Eui Ju ; Storm, Daniel R. / Oxidation of critical cysteine residues of type I adenylyl cyclase by o-iodosobenzoate or nitric oxide reversibly inhibits stimulation by calcium and calmodulin. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 10. pp. 7290-7296.
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AU - Storm, Daniel R.

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