Ligand-protein interactions in nitric oxide synthase

Denis L. Rousseau, David Li, Manon Couture, Syun Ru Yeh

Research output: Contribution to journalArticlepeer-review

94 Scopus citations


Nitric oxide synthases (NOSs) are heme proteins that catalyze the formation of nitric oxide (NO) from l-arginine and oxygen in a sequential two-step process. Three structurally similar isoforms have been identified that deliver NO to different tissues for specific functions. An understanding of the interactions of ligands with the protein is essential to determine the mechanism of catalysis, the design of inhibitors and the differential auto-inhibitory regulation of the enzymatic activity of the isoforms due to the binding of NO to the heme. Ligand-protein interactions in the three isoforms revealed by resonance Raman scattering studies are reviewed in this article. The CO-related modes in the CO-bound ferrous enzyme are sensitive to the presence of substrate, either l-arginine or N-hydroxy-l-arginine, in the distal pocket, but insensitive to the presence of the tetrahydrobiopterin (H4B) cofactor. In contrast, when NO is coordinated to the ferric heme, the NO is sensitive to the substrate only when H4B is present. Furthermore, in the NO-bound ferric enzyme, the addition of H4B induces a large heme distortion that may modulate heme reduction and thereby regulate the NO auto-inhibitory process. In the metastable O2-bound enzyme, l-arginine binding causes the appearance of a shoulder on the O-O stretching mode, suggesting a specific interaction of the heme-bound dioxygen with the bound-substrate that may be crucial for the oxygenation reaction of the substrate during the catalytic turn-over. It is postulated that spectroscopic differences in the oxy-complex are a consequence of the degree of protonation of the proximal cysteine ligand on the heme. Resonance Raman studies of NOSs expand our understanding of the mechanistic features of this important family of enzymes.

Original languageEnglish (US)
Pages (from-to)306-323
Number of pages18
JournalJournal of Inorganic Biochemistry
Issue number1
StatePublished - Jan 2005


  • Biophysics
  • Raman scattering
  • Vibrational spectroscopy

ASJC Scopus subject areas

  • Biochemistry
  • Inorganic Chemistry


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