Superoxide produced in the heme pocket of the β-chain of hemoglobin reacts with the β-93 cysteine to produce a thiyl radical

Academic Article


  • The role of the β-93 cysteine residue in the hemoglobin autoxidation process has been delineated by electron paramagnetic resonance. At low temperatures (8 K) after incubation at 235 K, free radical signals were detected. An analysis of the free radical spectrum produced implies that, besides the superoxide radical expected to be formed during autoxidation, an isotropic free radical is produced with a g(iso) of 2.0133. This g value is consistent with that expected for a sulfur radical. Blocking the β-93 sulfhydryl group with N-ethylmaleimide was found to eliminate the formation of the isotropic radical, but not the superoxide. This finding confirms the assignment of the isotropic radical as a thiyl radical originating from the oxidation of the cysteine SH group. A kinetic analysis of the time course for the formation of both the superoxide and thiyl radicals is consistent with a reversible electron transfer process between superoxide in the heme pocket of the β-chains and the cysteine residue. This reaction is expected to produce both a thiyl radical and a peroxide. Direct evidence for peroxide production comes from the detection of a transient Fe(III) heme peroxide complex. The significance of the electron transfer process producing a thiyl radical is discussed. It is shown that the formation of the thiyl radical decreases the rate of autoxidation for the β-chain and reduces heme degradation attributed to the reaction of superoxide with the heme. The insights gained from these low-temperature studies are believed to be relevant to room-temperature autoxidation.
  • Authors

    Published In

  • Biochemistry  Journal
  • Digital Object Identifier (doi)

    Pubmed Id

  • 4064428
  • Author List

  • Balagopalakrishna C; Abugo OO; Horsky J; Manoharan PT; Nagababu E; Rifkind JM
  • Start Page

  • 13194
  • End Page

  • 13202
  • Volume

  • 37
  • Issue

  • 38