Optical spectrum of myeloperoxidase. Origin of the red shift.
The optical spectrum of reduced myeloperoxidase (EC 1.11.1.7) displays an unusual red shift of the Soret band which is at 472 nm and the alpha-band which is at 636 nm. The spectral properties of myeloperoxidase can be modified by means of acid treatment. Upon short exposure to acid (pH 1.7) the red-shifted optical absorption spectrum of the reduced enzyme (lambda max at 472 nm) was blue-shifted (lambda max at 448 nm) but the spectrum of the reduced state could be restored by increasing the pH. By contrast, the resonance Raman spectra of both the oxidized and reduced enzyme are essentially the same at both pH 1.7 and pH 7.0. This shows that the optical spectrum and the resonance Raman spectrum are not directly correlated, which we interpret to indicate that the reversible effects of lower pH primarily affect the excited-state energy levels of the macrocycle. The EPR spectrum of the oxidized enzyme showed a reversible conversion from a high-spin rhombic spectrum (gx = 6.7, gy = 5.2) at neutral pH into a more axial high-spin spectrum (gx = gy = 5.8) at low pH. Upon prolonged exposure to acid (20 min) optical absorbance spectra, EPR spectra, resonance Raman spectra and the chlorinating activity were irreversibly affected. We propose that a negatively charged protonatable residue in the proximity of a pyrrole nucleus of the haem group is present that imposes the red shift in the optical absorption spectrum. This is consistent with the available X-ray structure data.
E.C. Slater Institute, University of Amsterdam, The Netherlands.
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R
Floris
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Kim
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Babcock
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Wever
D004578:Electron Spin Resonance Spectroscopy; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D007962:Leukocytes / Q000201:enzymology*; D010084:Oxidation-Reduction; D009195:Peroxidase / Q000097:blood / Q000737:chemistry*; D011487:Protein Conformation; D013053:Spectrophotometry / Q000379:methods; D013059:Spectrum Analysis, Raman; D014961:X-Ray Diffraction
D009195:Peroxidase