Electron transfer by neuronal nitric-oxide synthase is regulated by concerted interaction of calmodulin and two intrinsic regulatory elements

Linda J. Roman, Bettie Sue S Masters

Research output: Contribution to journalArticle

75 Citations (Scopus)

Abstract

The nitric-oxide synthases (NOSs) are modular, cofactor-containing enzymes, divided into a heme-containing oxygenase domain and an FMN- and FAD-containing reductase domain. The domains are connected by a calmodulin (CaM)-binding sequence, occupancy of which is required for nitric oxide (NO) production. Two additional CaM-modulated regulatory elements are present in the reductase domains of the constitutive isoforms, the autoregulatory region (AR) and the C-terminal tail region. Deletion of the AR reduces CaM stimulation of electron flow through the reductase domain from 10-fold in wild-type nNOS to 2-fold in the mutant. Deletion of the C terminus yields an enzyme with greatly enhanced reductase activity in the absence of CaM but with activity equivalent to that of wild-type enzyme in its presence. A mutant in which both the AR and C terminus were deleted completely loses CaM modulation through the reductase domain. Thus, transduction of the CaM effect through the reductase domain of nNOS is dependent on these elements. Formation of nitric oxide is, however, still stimulated by CaM in all three mutants. A CaM molecule in which the N-terminal lobe was replaced by the C-terminal lobe (CaMCC) supported NO synthesis by the deletion mutants but not by wild-type nNOS. We propose a model in which the AR, the C-terminal tail, and CaM interact directly to regulate the conformational state of the reductase domain of nNOS.

Original languageEnglish (US)
Pages (from-to)23111-23118
Number of pages8
JournalJournal of Biological Chemistry
Volume281
Issue number32
DOIs
StatePublished - Aug 11 2006

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Nitric Oxide Synthase Type I
Calmodulin
Electrons
Oxidoreductases
Nitric Oxide
Enzymes
Flavin Mononucleotide
Heme Oxygenase (Decyclizing)
Oxygenases
Flavin-Adenine Dinucleotide
Coenzymes
Heme
Nitric Oxide Synthase
Protein Isoforms
Modulation
Molecules

ASJC Scopus subject areas

  • Biochemistry

Cite this

Electron transfer by neuronal nitric-oxide synthase is regulated by concerted interaction of calmodulin and two intrinsic regulatory elements. / Roman, Linda J.; Masters, Bettie Sue S.

In: Journal of Biological Chemistry, Vol. 281, No. 32, 11.08.2006, p. 23111-23118.

Research output: Contribution to journalArticle

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