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Biophysical journal2002; 83(5); 2845-2855; doi: 10.1016/S0006-3495(02)75293-4

Comparative study of tyrosine radicals in hemoglobin and myoglobins treated with hydrogen peroxide.

Abstract: The reactions of hydrogen peroxide with human methemoglobin, sperm whale metmyoglobin, and horse heart metmyoglobin were studied by electron paramagnetic resonance (EPR) spectroscopy at 10 K and room temperature. The singlet EPR signal, one of the three signals seen in these systems at 10 K, is characterized by a poorly resolved, but still detectable, hyperfine structure that can be used to assign it to a tyrosyl radical. The singlet is detectable as a quintet at room temperature in methemoglobin with identical spectral features to those of the well characterized tyrosyl radical in photosystem II. Hyperfine splitting constants found for Tyr radicals were used to find the rotation angle of the phenoxyl group. Analysis of these angles in the crystal structures suggests that the radical resides on Tyr151 in sperm whale myoglobin, Tyr133 in soybean leghemoglobin, and either alphaTyr42, betaTyr35, or betaTyr130 in hemoglobin. In the sperm whale metmyoglobin Tyr103Phe mutant, there is no detectable tyrosyl radical present. Yet the rotation angle of Tyr103 (134 degrees) is too large to account for the observed EPR spectrum in the wild type. Tyr103 is the closest to the heme. We suggest that Tyr103 is the initial site of the radical, which then rapidly migrates to Tyr151.
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Publication Date: 2002-11-05 PubMed ID: 12414716PubMed Central: PMC1302368DOI: 10.1016/S0006-3495(02)75293-4Google Scholar: Lookup
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  • Comparative Study
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research article examines how hydrogen peroxide interacts with different types of hemoglobin and myoglobins, commonly found in the body, by using electron paramagnetic resonance (EPR) spectroscopy. The researchers demonstrate the complex formation and movement of tyrosine radicals in these proteins.

Methodology

  • The researchers studied the reactions of hydrogen peroxide with three different hemoglobin and myoglobin proteins: human methemoglobin, sperm whale metmyoglobin, and horse heart metmyoglobin. They chose these proteins to represent a range of hemoglobins and myoglobins.
  • The reactions were further examined at two temperatures: 10 K (-263.15 degrees Celsius) and room temperature.
  • Using electron paramagnetic resonance (EPR) spectroscopy, a technique that identifies species with unpaired electrons (like radicals), the scientists were able to look closely at the reactions between the proteins and hydrogen peroxide.

Findings

  • The scientists identified a distinct EPR signal which they attributed to a tyrosyl (tyrosine) radical, a reactive molecule found in proteins.
  • This singlet EPR signal indicated a tyrosyl radical present in the proteins at 10 degrees Kelvin and at room temperature.
  • They found that the radical’s signal matches the known characteristics of a tyrosyl radical present in photosystem II, a part of light-dependent reactions in photosynthesis, reinforcing the identification of the radical as a tyrosyl radical.
  • In further investigations, they used the spectral characteristics of these tyrosyl radicals to determine the rotation angle of the phenoxyl group, a substructure commonly found in organic molecules.

Location of the tyrosyl radical

  • Their data analysis suggested that the tyrosyl radical sits at a specific location (Tyr103 initially morphing into Tyr151) on sperm whale metmyoglobin. This movement from Tyr103 to Tyr151 suggests a migration of the radical within the protein structure.
  • In the soybean leghemoglobin, the radical was predicted to reside at Tyr133 position.
  • In the case of human hemoglobin, this radical could be on either alphaTyr42, betaTyr35, or betaTyr130.

Implications

  • The research provides valuable insight into the chemistry and biochemistry of these proteins, their reaction patterns, and the characteristics of the tyrosyl radicals. This can be helpful for understanding disease mechanisms possibly related to oxidative stress where reactive species like radicals play a crucial role.

Cite This Article

APA
Svistunenko DA, Dunne J, Fryer M, Nicholls P, Reeder BJ, Wilson MT, Bigotti MG, Cutruzzolà F, Cooper CE. (2002). Comparative study of tyrosine radicals in hemoglobin and myoglobins treated with hydrogen peroxide. Biophys J, 83(5), 2845-2855. https://doi.org/10.1016/S0006-3495(02)75293-4

Publication

Biophysical journal
ISSN: 0006-3495
NlmUniqueID: 0370626
Country: United States
Language: English
Volume: 83
Issue: 5
Pages: 2845-2855

Researcher Affiliations

Svistunenko, Dimitri A
  • Department of Biological Sciences, Central Campus, University of Essex, Wivenhoe Park, Colchester CO4 3SQ, UK. svist@essex.ac.uk
Dunne, Jacqueline
    Fryer, Michael
      Nicholls, Peter
        Reeder, Brandon J
          Wilson, Michael T
            Bigotti, Maria Giulia
              Cutruzzolà, Francesca
                Cooper, Chris E

                  MeSH Terms

                  • Animals
                  • Arabidopsis
                  • Electron Spin Resonance Spectroscopy
                  • Free Radicals
                  • Hemoglobins / chemistry
                  • Horses
                  • Humans
                  • Hydrogen Peroxide / pharmacology
                  • Iron
                  • Myoglobin / chemistry
                  • Photosynthesis
                  • Protein Conformation
                  • Temperature
                  • Tyrosine / chemistry
                  • Whales

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