FeNO structure in distal pocket mutants of myoglobin based on resonance Raman spectroscopy

Candace M. Coyle, Kathleen M. Vogel, Thomas S. Rush, Pawel M. Kozlowski, Robert Williams, Thomas G. Spiro, Yi Dou, Masao Ikeda-Saito, John S. Olson, Marek Z. Zgierski

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73 Scopus citations


FeNO vibrational frequencies were investigated for a series of myoglobin mutants using isotope-edited resonance Raman spectra of 15/14NO adducts, which reveal the FeNO and NO stretching modes. The latter give rise to doublet bands, as a result of Fermi resonances with coincident porphyrin vibrations; these doublets were analyzed by curve-fitting to obtain the vNO frequencies. Variations in vNO among the mutants correlate with the reported vCO variations for the CO adducts of the same mutants. The correlation has a slope near unity, indicating equal sensitivity of the NO and CO bonds to polar influences in the heme pocket. A few mutants deviate from the correlation, indicating that distal interactions differ for the NO and CO adducts, probably because of the differing distal residue geometries. In contrast to the strong and consistent vFeC/vCO correlation found for the CO adducts, vFeN correlates only weakly with vNO, and the slope of the correlation depends on which residue is being mutated. This variability is suggested to arise from steric interactions, which change the FeNO angle and therefore alter the Fe-NO and N-O bond orders. This effect is modeled with Density Functional Theory (DFT) and is rationalized on the basis of a valence isomer bonding model. The FeNO unit, which is naturally bent, is a more sensitive reporter of steric interactions than the FeCO unit, which is naturally linear. An important additional factor is the strength of the bond to the proximal ligand, which modulates the valence isomer equilibrium. The FeNO unit is bent more strongly in MbNO than in protein-free heme-NO complexes because of a combination of a strengthened proximal bond and distal interactions.

Original languageEnglish (US)
Pages (from-to)4896-4903
Number of pages8
Issue number17
StatePublished - May 6 2003
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry


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