X-ray absorption and resonance raman spectroscopy of human myeloperoxidase at neutral and acid pH

K. T. Yue, K. L. Taylor, J. M. Kinkade, R. B. Sinclair, Linda S Powers

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Myeloperoxidase (MPG), an important enzyme in the oxygen-dependent host defense system of human polymorphonuclear leukocytes, utilizes hydrogen peroxide to catalyze the production of hypochlorous acid, an oxidizing bactericidal agent. While MPO shows significant sequence homology with other peroxidases and this homology is particularly striking among the active-site residues, MPO exhibits unusual spectral features and the unique ability to catalyze the oxidation of chloride ions. We have investigated the MPO active-site with X-ray absorption (XAS) and resonance Raman (RRS) spectroscopies at neutral pH and also at the physiological acidic pH (pH ~ 3) and have compared these results with those of horseradish peroxidase (HRP). At pH 7.5, XAS results show that the iron heme active site is 6-coordinate where the distal ligand is likely nitrogen or oxygen, but not sulfur. The heme is distorted compared to HRP, other peroxidases, and heme compounds, but at pH ~ 3, the distal ligand is lost and the heme is less distorted. RRS results under identical pH conditions show that the skeletal core-size sensitive modes and ν3, are shifted to higher frequency at pH ~ 3 indicating a 6- to 5-coordination change of high spin ferric heme. In addition, a new band at 270 cm-1 is observed at pH ~ 3 which is consistent with the loss of the sixth ligand. The higher symmetry of the heme at pH ~ 3 is reflected by a single ν4 in the (RRS) spectrum. HRP also loses its loosely associated distal water at this pH, but little change in heme distortion is observed. This change suggests that loss of the distal ligand in MPO releases stress on the heme which may facilitate binding of chloride ion.

Original languageEnglish (US)
Pages (from-to)282-294
Number of pages13
JournalBiochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
Volume1338
Issue number2
DOIs
StatePublished - Apr 4 1997
Externally publishedYes

Fingerprint

Raman Spectrum Analysis
X ray absorption
Heme
Peroxidase
Raman spectroscopy
X-Rays
Acids
Horseradish Peroxidase
Ligands
Peroxidases
Catalytic Domain
Chlorides
Ions
Oxygen
Hypochlorous Acid
Sulfur
Hydrogen Peroxide
Sequence Homology
Nitrogen
Iron

Keywords

  • Active heme site
  • Myeloperoxidase
  • Peroxidase
  • pH effect

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Structural Biology
  • Biophysics

Cite this

X-ray absorption and resonance raman spectroscopy of human myeloperoxidase at neutral and acid pH. / Yue, K. T.; Taylor, K. L.; Kinkade, J. M.; Sinclair, R. B.; Powers, Linda S.

In: Biochimica et Biophysica Acta (BBA)/Protein Structure and Molecular, Vol. 1338, No. 2, 04.04.1997, p. 282-294.

Research output: Contribution to journalArticle

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