Heme coordination and structure of the catalytic site in nitric oxide synthase

J. Wang, D. J. Stuehr, M. Ikeda-Saito, Denis L. Rousseau

Research output: Contribution to journalArticle

91 Citations (Scopus)

Abstract

Nitric oxide (NO), recently found to play many physiological roles, is generated by the catalysis of L-arginine and O2 to L-citrulline and NO by nitric oxide synthases (NOSs). Resonance Raman spectra from the heme of resting, reduced, and CO-bound forms of rat brain NOS firmly establish that the enzyme belongs to the P-450 class of enzymes. The electron density marker line (v4) in the Raman spectrum of ligand-free ferrous NOS has a low frequency (1347 cm-1), indicating a thiolate axial ligand on the heme. The assignment of a thiolate axial ligand is confirmed in the CO-bound form of the enzyme by the frequency of the Fe-C-O bending mode at 562 cm-1. The heme in resting NOS is five-coordinate high spin and thereby differs from the resting state of most substrate-free P450s, which are predominantly six- coordinate low spin. The frequency of the Fe-CO stretching mode in the CO- bound enzyme at 491 cm-1, identified by isotope substitution, is higher than that in substrate-free P-450s. Thus, in the ferric and the CO-bound forms of the enzyme, the sixth-ligand binding site on the heme is restricted by steric or hydrophobic interactions. In addition, the Fe-CO stretching mode is broad (30 cm-1) and may be resolved into two overlapping lines of equal intensity, indicating that the heme domains can adopt two distinct conformations.

Original languageEnglish (US)
Pages (from-to)22255-22258
Number of pages4
JournalJournal of Biological Chemistry
Volume268
Issue number30
StatePublished - 1993
Externally publishedYes

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Carbon Monoxide
Heme
Nitric Oxide Synthase
Catalytic Domain
Ligands
Enzymes
Stretching
Raman scattering
Nitric Oxide
Citrulline
Substrates
Catalysis
Hydrophobic and Hydrophilic Interactions
Isotopes
Cytochrome P-450 Enzyme System
Carrier concentration
Arginine
Conformations
Substitution reactions
Binding Sites

ASJC Scopus subject areas

  • Biochemistry

Cite this

Heme coordination and structure of the catalytic site in nitric oxide synthase. / Wang, J.; Stuehr, D. J.; Ikeda-Saito, M.; Rousseau, Denis L.

In: Journal of Biological Chemistry, Vol. 268, No. 30, 1993, p. 22255-22258.

Research output: Contribution to journalArticle

Wang, J. ; Stuehr, D. J. ; Ikeda-Saito, M. ; Rousseau, Denis L. / Heme coordination and structure of the catalytic site in nitric oxide synthase. In: Journal of Biological Chemistry. 1993 ; Vol. 268, No. 30. pp. 22255-22258.
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