Skip to main content
Biophysical Journal logoLink to Biophysical Journal
. 2000 Apr;78(4):2107–2115. doi: 10.1016/S0006-3495(00)76757-9

Proton electron nuclear double resonance from nitrosyl horse heart myoglobin: the role of His-E7 and Val-E11.

M Flores 1, E Wajnberg 1, G Bemski 1
PMCID: PMC1300802  PMID: 10733988

Abstract

Electron nuclear double resonance (ENDOR) spectroscopy has been used to study protons in nitrosyl horse heart myoglobin (MbNO). (1)H ENDOR spectra were recorded for different settings of the magnetic field. Detailed analysis of the ENDOR powder spectra, using computer simulation, based on the "orientation-selection" principle, leads to the identification of the available protons in the heme pocket. We observe hyperfine interactions of the N(HisF8)-Fe(2+)-N(NO) complex with five protons in axial and with eight protons in the rhombic symmetry along different orientations, including those of the principal axes of the g-tensor. Protons from His-E7 and Val-E11 residues are identified in the two symmetries, rhombic and axial, exhibited by MbNO. Our results indicate that both residues are present inside the heme pocket and help to stabilize one particular conformation.

Full Text

The Full Text of this article is available as a PDF (195.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Braunstein D. P., Chu K., Egeberg K. D., Frauenfelder H., Mourant J. R., Nienhaus G. U., Ormos P., Sligar S. G., Springer B. A., Young R. D. Ligand binding to heme proteins: III. FTIR studies of His-E7 and Val-E11 mutants of carbonmonoxymyoglobin. Biophys J. 1993 Dec;65(6):2447–2454. doi: 10.1016/S0006-3495(93)81310-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Feher G., Isaacson R. A., Scholes C. P., Nagel R. Electron nuclear double resonance (ENDOR) investigation on myoglobin and hemoglobin. Ann N Y Acad Sci. 1973 Dec 31;222:86–101. doi: 10.1111/j.1749-6632.1973.tb15254.x. [DOI] [PubMed] [Google Scholar]
  3. Flores M., Wajnberg E., Bemski G. Temperature dependence of Q-band electron paramagnetic resonance spectra of nitrosyl heme proteins. Biophys J. 1997 Dec;73(6):3225–3229. doi: 10.1016/S0006-3495(97)78347-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Frauenfelder H., Sligar S. G., Wolynes P. G. The energy landscapes and motions of proteins. Science. 1991 Dec 13;254(5038):1598–1603. doi: 10.1126/science.1749933. [DOI] [PubMed] [Google Scholar]
  5. Hanson J. C., Schoenborn B. P. Real space refinement of neutron diffraction data from sperm whale carbonmonoxymyoglobin. J Mol Biol. 1981 Nov 25;153(1):117–146. doi: 10.1016/0022-2836(81)90530-1. [DOI] [PubMed] [Google Scholar]
  6. Hong M. K., Braunstein D., Cowen B. R., Frauenfelder H., Iben I. E., Mourant J. R., Ormos P., Scholl R., Schulte A., Steinbach P. J. Conformational substates and motions in myoglobin. External influences on structure and dynamics. Biophys J. 1990 Aug;58(2):429–436. doi: 10.1016/S0006-3495(90)82388-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hori H., Ikeda-Saito M., Yonetani T. Single crystal EPR of myoglobin nitroxide. Freezing-induced reversible changes in the molecular orientation of the ligand. J Biol Chem. 1981 Aug 10;256(15):7849–7855. [PubMed] [Google Scholar]
  8. Jewsbury P., Kitagawa T. The distal residue-CO interaction in carbonmonoxy myoglobins: a molecular dynamics study of two distal histidine tautomers. Biophys J. 1994 Dec;67(6):2236–2250. doi: 10.1016/S0006-3495(94)80708-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Li T., Quillin M. L., Phillips G. N., Jr, Olson J. S. Structural determinants of the stretching frequency of CO bound to myoglobin. Biochemistry. 1994 Feb 15;33(6):1433–1446. doi: 10.1021/bi00172a021. [DOI] [PubMed] [Google Scholar]
  10. Lim M., Jackson T. A., Anfinrud P. A. Binding of CO to myoglobin from a heme pocket docking site to form nearly linear Fe-C-O. Science. 1995 Aug 18;269(5226):962–966. doi: 10.1126/science.7638619. [DOI] [PubMed] [Google Scholar]
  11. Mathews A. J., Rohlfs R. J., Olson J. S., Tame J., Renaud J. P., Nagai K. The effects of E7 and E11 mutations on the kinetics of ligand binding to R state human hemoglobin. J Biol Chem. 1989 Oct 5;264(28):16573–16583. [PubMed] [Google Scholar]
  12. Olson J. S., Mathews A. J., Rohlfs R. J., Springer B. A., Egeberg K. D., Sligar S. G., Tame J., Renaud J. P., Nagai K. The role of the distal histidine in myoglobin and haemoglobin. Nature. 1988 Nov 17;336(6196):265–266. doi: 10.1038/336265a0. [DOI] [PubMed] [Google Scholar]
  13. Peng S. M., Ibers J. A. Stereochemistry of carbonylmetalloporphyrins. The structure of (pyridine)(carbonyl)(5, 10, 15, 20-tetraphenylprophinato)iron(II). J Am Chem Soc. 1976 Dec 8;98(25):8032–8036. doi: 10.1021/ja00441a025. [DOI] [PubMed] [Google Scholar]
  14. Perutz M. F. Myoglobin and haemoglobin: role of distal residues in reactions with haem ligands. Trends Biochem Sci. 1989 Feb;14(2):42–44. doi: 10.1016/0968-0004(89)90039-x. [DOI] [PubMed] [Google Scholar]
  15. Perutz M. F. Stereochemistry of cooperative effects in haemoglobin. Nature. 1970 Nov 21;228(5273):726–739. doi: 10.1038/228726a0. [DOI] [PubMed] [Google Scholar]
  16. Phillips S. E., Schoenborn B. P. Neutron diffraction reveals oxygen-histidine hydrogen bond in oxymyoglobin. Nature. 1981 Jul 2;292(5818):81–82. doi: 10.1038/292081a0. [DOI] [PubMed] [Google Scholar]
  17. Phillips S. E. Structure and refinement of oxymyoglobin at 1.6 A resolution. J Mol Biol. 1980 Oct 5;142(4):531–554. doi: 10.1016/0022-2836(80)90262-4. [DOI] [PubMed] [Google Scholar]
  18. Rohlfs R. J., Mathews A. J., Carver T. E., Olson J. S., Springer B. A., Egeberg K. D., Sligar S. G. The effects of amino acid substitution at position E7 (residue 64) on the kinetics of ligand binding to sperm whale myoglobin. J Biol Chem. 1990 Feb 25;265(6):3168–3176. [PubMed] [Google Scholar]
  19. Shaanan B. Structure of human oxyhaemoglobin at 2.1 A resolution. J Mol Biol. 1983 Nov 25;171(1):31–59. doi: 10.1016/s0022-2836(83)80313-1. [DOI] [PubMed] [Google Scholar]
  20. Springer B. A., Egeberg K. D., Sligar S. G., Rohlfs R. J., Mathews A. J., Olson J. S. Discrimination between oxygen and carbon monoxide and inhibition of autooxidation by myoglobin. Site-directed mutagenesis of the distal histidine. J Biol Chem. 1989 Feb 25;264(6):3057–3060. [PubMed] [Google Scholar]
  21. Tierney D. L., Huang H., Martasek P., Masters B. S., Silverman R. B., Hoffman B. M. ENDOR spectroscopic evidence for the position and structure of NG-hydroxy-L-arginine bound to holo-neuronal nitric oxide synthase. Biochemistry. 1999 Mar 23;38(12):3704–3710. doi: 10.1021/bi982904r. [DOI] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES