Assignment of heme methyl 1H-NMR resonances of high-spin and low-spin ferric complexes of cytochrome p450cam using one-dimensional and two-dimensional paramagnetic signals enhancement (PASE) magnetization transfer experiments

Eur J Biochem. 2000 Jan;267(1):216-21. doi: 10.1046/j.1432-1327.2000.00995.x.

Abstract

An 1H-NMR study of ferric cytochrome P450cam in different paramagnetic states was performed. Assignment of three heme methyl resonances of the isocyanide adduct of cytochrome P450 in the ferric low-spin state was recently performed using electron exchange in the presence of putidaredoxin [Mouro, C., Bondon, A., Jung, C., Hui Bon Hoa, G., De Certaines, J.D., Spencer, R.G.S. & Simonneaux, G. (1999) FEBS Lett. 455, 302-306]. In this study, heme methyl protons of cytochrome P450 in the native high-spin and low-spin states were assigned through one-dimensional and two-dimensional magnetization transfer spectroscopy using the paramagnetic signals enhancement (PASE) method. The order of the methyl proton chemical shifts is inverted between high-spin and low-spin states. The methyl order observed in the ferric low-spin isocyanide complexes is related to the orientation of the cysteinate ligand.

MeSH terms

  • Camphor / metabolism
  • Camphor 5-Monooxygenase / chemistry*
  • Camphor 5-Monooxygenase / metabolism*
  • Electron Spin Resonance Spectroscopy
  • Ferredoxins / metabolism
  • Ferric Compounds / metabolism*
  • Heme / chemistry*
  • Heme / metabolism*
  • Ligands
  • Nitriles / metabolism
  • Nuclear Magnetic Resonance, Biomolecular*
  • Protons

Substances

  • Ferredoxins
  • Ferric Compounds
  • Ligands
  • Nitriles
  • Protons
  • butyl isocyanide
  • Heme
  • putidaredoxin
  • ethyl isocyanide
  • Camphor
  • Camphor 5-Monooxygenase