title: 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.
authors.affiliation: Department of Biological Sciences, Central Campus, University of Essex, Wivenhoe Park, Colchester CO4 3SQ, UK. svist@essex.ac.uk
authors.forename: Dimitri A
authors.initials: DA
authors.lastname: Svistunenko
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Dunne
authors.forename: Michael
authors.initials: M
authors.lastname: Fryer
authors.forename: Peter
authors.initials: P
authors.lastname: Nicholls
authors.forename: Brandon J
authors.initials: BJ
authors.lastname: Reeder
authors.forename: Michael T
authors.initials: MT
authors.lastname: Wilson
authors.forename: Maria Giulia
authors.initials: MG
authors.lastname: Bigotti
authors.forename: Francesca
authors.initials: F
authors.lastname: Cutruzzolà
authors.forename: Chris E
authors.initials: CE
authors.lastname: Cooper
mesh_terms: D000818:Animals; D017360:Arabidopsis; D004578:Electron Spin Resonance Spectroscopy; D005609:Free Radicals; D006454:Hemoglobins / Q000737:chemistry*; D006736:Horses; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D007501:Iron; D009211:Myoglobin / Q000737:chemistry*; D010788:Photosynthesis; D011487:Protein Conformation; D013696:Temperature; D014443:Tyrosine / Q000737:chemistry*; D014907:Whales
chemical_list: D005609:Free Radicals; D006454:Hemoglobins; D009211:Myoglobin; D014443:Tyrosine; D006861:Hydrogen Peroxide; D007501:Iron