Heme coordination of NO in NO synthase

Jianling Wang, Denis L. Rousseau, Husam M. Abu-Soud, Dennis J. Stuehr

Research output: Contribution to journalArticle

123 Citations (Scopus)

Abstract

A current question in nitric oxide (NO) biology is whether NO can act as a feedback inhibitor of NO synthase (NOS). We have approached this problem by examining the interaction of NO with neuronal NOS by optical absorption and resonance Raman scattering spectroscopies. Under an inert atmosphere NO coordinated to the heme iron in both the oxidized and reduced forms of NOS. The Soret and visible optical absorption transitions are detected at 436 and at 567 nm, respectively, in the Fe2+-NO heme complex and at 440 nm and at 549 and 580 nm, respectively, in the Fe3+-NO heme complex. In the resonance Raman spectrum of the ferrous complex the Fe-NO stretching mode is located at 549 cm-1 in the presence of L-arginine and at 536 cm-1 in the absence of L-arginine, whereas in the ferric enzyme the mode is located at 540 cm-1 (in the absence of L-arginine). The interaction between bound L-arginine and the NO indicates that L-arginine binds directly over the heme just as do the substrates in cytochrome P-450s. In the absence of L-arginine, NO readily oxidized the ferrous heme iron. The oxidation was prevented by the presence of bound L-arginine and enabled NOS to form a stable ferrous NO complex. Under oxygen-limited conditions, NO generated by neuronal NOS coordinated to its heme iron and formed a spectrally detectable ferrous-NO complex. Taken together, our results show that NO can bind to both ferric and ferrous NOS and may inhibit NO synthesis through its binding to the heme iron during catalysis.

Original languageEnglish (US)
Pages (from-to)10512-10516
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume91
Issue number22
DOIs
StatePublished - Oct 25 1994
Externally publishedYes

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Heme
Nitric Oxide Synthase
Nitric Oxide
Arginine
Iron
Raman Spectrum Analysis
Cytochromes
Catalysis
Atmosphere

Keywords

  • cytochrome P-450
  • flavoprotein
  • L-arginine
  • Raman scattering

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Heme coordination of NO in NO synthase. / Wang, Jianling; Rousseau, Denis L.; Abu-Soud, Husam M.; Stuehr, Dennis J.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 91, No. 22, 25.10.1994, p. 10512-10516.

Research output: Contribution to journalArticle

Wang, Jianling ; Rousseau, Denis L. ; Abu-Soud, Husam M. ; Stuehr, Dennis J. / Heme coordination of NO in NO synthase. In: Proceedings of the National Academy of Sciences of the United States of America. 1994 ; Vol. 91, No. 22. pp. 10512-10516.
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AB - A current question in nitric oxide (NO) biology is whether NO can act as a feedback inhibitor of NO synthase (NOS). We have approached this problem by examining the interaction of NO with neuronal NOS by optical absorption and resonance Raman scattering spectroscopies. Under an inert atmosphere NO coordinated to the heme iron in both the oxidized and reduced forms of NOS. The Soret and visible optical absorption transitions are detected at 436 and at 567 nm, respectively, in the Fe2+-NO heme complex and at 440 nm and at 549 and 580 nm, respectively, in the Fe3+-NO heme complex. In the resonance Raman spectrum of the ferrous complex the Fe-NO stretching mode is located at 549 cm-1 in the presence of L-arginine and at 536 cm-1 in the absence of L-arginine, whereas in the ferric enzyme the mode is located at 540 cm-1 (in the absence of L-arginine). The interaction between bound L-arginine and the NO indicates that L-arginine binds directly over the heme just as do the substrates in cytochrome P-450s. In the absence of L-arginine, NO readily oxidized the ferrous heme iron. The oxidation was prevented by the presence of bound L-arginine and enabled NOS to form a stable ferrous NO complex. Under oxygen-limited conditions, NO generated by neuronal NOS coordinated to its heme iron and formed a spectrally detectable ferrous-NO complex. Taken together, our results show that NO can bind to both ferric and ferrous NOS and may inhibit NO synthesis through its binding to the heme iron during catalysis.

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