Heme protein radicals

Formation, fate, and biological consequences

Cecilia R Giulivi, Enrique Cadenas

Research output: Contribution to journalArticle

121 Citations (Scopus)

Abstract

The oxidation of myoglobin by H2O2 yields ferrylmyoglobin, which contains two oxidizing equivalents, the oxoferryl complex and an amino acid radical. This study examines the electron paramagnetic resonance (EPR) properties of the resulting amino acid radicals and their inherent kinetic features at [H2O2]/[protein] ratios close to physiological conditions (i.e., ≤ 1). The EPR spectrum obtained with continuous flow at room temperature consisted of a composite of three signals, a low intensity signal and two high intensity signals. The former had a g-value of 2 014, contributed 10-15% to the overall spectrum and was ascribed to a peroxyl radical. Of the two high intensity signals, one consisted of a six-line spectrum (g = 2.0048) that contributed approximately 17-19% to the overall signal; hyperfine splitting constants to ring protons permitted to identify this signal as a tyrosyl radical. The other high intensity signal (with similar g-value and underlying that of the tyrosyl radical) was ascribed to an aromatic amino acid upon comparison with the EPR characteristics for radicals in aromatic amino acid-containing peptides Analysis of these data in connection with amino acid analysis and the EPR spectra obtained under similar conditions with another hemoprotein, hemoglobin, allowed to suggest a mechanism for the formation of the protein radicals in myoglobin. The aromatic amino acid radical was observed to be relatively long lived in close proximity to the heme iron. Hence, it is likely that this is the first site of protein radical; reduction of the oxoferryl complex by Tyr (Fe(IV)=O + Tyr-OH + H+ → Fe(III) + H2O + Tyr-O·)-and alternatively by other amino acids-leads to the subsequent formation of other amino acid radicals within an electron-transfer process throughout the protein. This view suggests that the protein radical(s) is highly delocalized within the globin moiety in a dynamic process encompassing electron tunneling through the backbone chain or H-bonds and leading to the formation of secondary radicals.

Original languageEnglish (US)
Pages (from-to)269-279
Number of pages11
JournalFree Radical Biology and Medicine
Volume24
Issue number2
DOIs
StatePublished - Jan 15 1998
Externally publishedYes

Fingerprint

Hemeproteins
Electron Spin Resonance Spectroscopy
Aromatic Amino Acids
Paramagnetic resonance
Amino Acids
Myoglobin
Proteins
Electrons
Electron tunneling
Globins
Heme
Protons
Hemoglobins
Iron
Oxidation
Peptides
Kinetics
Temperature
Composite materials

Keywords

  • Electron paramagnetic resonance
  • Electron tunneling
  • Hemoglobin
  • Hydrogen peroxide
  • Myoglobin
  • Myoglobin radicals
  • Oxoferryl
  • Protein radicals
  • Tyrosyl radicals

ASJC Scopus subject areas

  • Medicine(all)
  • Toxicology
  • Clinical Biochemistry

Cite this

Heme protein radicals : Formation, fate, and biological consequences. / Giulivi, Cecilia R; Cadenas, Enrique.

In: Free Radical Biology and Medicine, Vol. 24, No. 2, 15.01.1998, p. 269-279.

Research output: Contribution to journalArticle

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N2 - The oxidation of myoglobin by H2O2 yields ferrylmyoglobin, which contains two oxidizing equivalents, the oxoferryl complex and an amino acid radical. This study examines the electron paramagnetic resonance (EPR) properties of the resulting amino acid radicals and their inherent kinetic features at [H2O2]/[protein] ratios close to physiological conditions (i.e., ≤ 1). The EPR spectrum obtained with continuous flow at room temperature consisted of a composite of three signals, a low intensity signal and two high intensity signals. The former had a g-value of 2 014, contributed 10-15% to the overall spectrum and was ascribed to a peroxyl radical. Of the two high intensity signals, one consisted of a six-line spectrum (g = 2.0048) that contributed approximately 17-19% to the overall signal; hyperfine splitting constants to ring protons permitted to identify this signal as a tyrosyl radical. The other high intensity signal (with similar g-value and underlying that of the tyrosyl radical) was ascribed to an aromatic amino acid upon comparison with the EPR characteristics for radicals in aromatic amino acid-containing peptides Analysis of these data in connection with amino acid analysis and the EPR spectra obtained under similar conditions with another hemoprotein, hemoglobin, allowed to suggest a mechanism for the formation of the protein radicals in myoglobin. The aromatic amino acid radical was observed to be relatively long lived in close proximity to the heme iron. Hence, it is likely that this is the first site of protein radical; reduction of the oxoferryl complex by Tyr (Fe(IV)=O + Tyr-OH + H+ → Fe(III) + H2O + Tyr-O·)-and alternatively by other amino acids-leads to the subsequent formation of other amino acid radicals within an electron-transfer process throughout the protein. This view suggests that the protein radical(s) is highly delocalized within the globin moiety in a dynamic process encompassing electron tunneling through the backbone chain or H-bonds and leading to the formation of secondary radicals.

AB - The oxidation of myoglobin by H2O2 yields ferrylmyoglobin, which contains two oxidizing equivalents, the oxoferryl complex and an amino acid radical. This study examines the electron paramagnetic resonance (EPR) properties of the resulting amino acid radicals and their inherent kinetic features at [H2O2]/[protein] ratios close to physiological conditions (i.e., ≤ 1). The EPR spectrum obtained with continuous flow at room temperature consisted of a composite of three signals, a low intensity signal and two high intensity signals. The former had a g-value of 2 014, contributed 10-15% to the overall spectrum and was ascribed to a peroxyl radical. Of the two high intensity signals, one consisted of a six-line spectrum (g = 2.0048) that contributed approximately 17-19% to the overall signal; hyperfine splitting constants to ring protons permitted to identify this signal as a tyrosyl radical. The other high intensity signal (with similar g-value and underlying that of the tyrosyl radical) was ascribed to an aromatic amino acid upon comparison with the EPR characteristics for radicals in aromatic amino acid-containing peptides Analysis of these data in connection with amino acid analysis and the EPR spectra obtained under similar conditions with another hemoprotein, hemoglobin, allowed to suggest a mechanism for the formation of the protein radicals in myoglobin. The aromatic amino acid radical was observed to be relatively long lived in close proximity to the heme iron. Hence, it is likely that this is the first site of protein radical; reduction of the oxoferryl complex by Tyr (Fe(IV)=O + Tyr-OH + H+ → Fe(III) + H2O + Tyr-O·)-and alternatively by other amino acids-leads to the subsequent formation of other amino acid radicals within an electron-transfer process throughout the protein. This view suggests that the protein radical(s) is highly delocalized within the globin moiety in a dynamic process encompassing electron tunneling through the backbone chain or H-bonds and leading to the formation of secondary radicals.

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