Identification of a nickel(II) binding site on hemoglobin which confers susceptibility to oxidative deamination and intramolecular cross-linking

J Biol Chem. 1998 May 22;273(21):13037-46. doi: 10.1074/jbc.273.21.13037.

Abstract

Complexation of Ni(II) with native state recombinant hemoglobin is shown to produce NH2-terminal deamination and globin cross-linking in the presence of the oxidant potassium peroxymonosulfate (OxoneTM). Both the oxidative deamination and cross-linking are exclusive to the beta chains. Recombinant hemoglobin mutants have been created to identify protein sequence requirements for these reactions. It was found that His-2 of the beta globin is required for redox active Ni(II) complexation, oxidative deamination, and cross-linking. The oxidative deamination results in the formation of a free carbonyl in place of the NH2-terminal amine of the beta chain. Most cross-linking of the beta globin occurs intramolecularly, forming beta globin dimers. Structural characterization of the beta globin dimers indicates the presence of heterogeneous cross-links within the central hemoglobin cavity between the NH2 terminus of one beta chain and the COOH-terminal region of the other.

MeSH terms

  • Amino Acid Sequence
  • Binding Sites
  • Chromatography, High Pressure Liquid
  • Deamination
  • Dimerization
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / genetics
  • Hemoglobins / chemistry
  • Hemoglobins / genetics
  • Hemoglobins / metabolism*
  • Humans
  • Mass Spectrometry
  • Molecular Sequence Data
  • Nickel / metabolism*
  • Oxidation-Reduction
  • Peptide Mapping
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Sulfuric Acids / chemistry

Substances

  • Hemoglobins
  • Recombinant Proteins
  • Sulfuric Acids
  • Nickel
  • potassium peroxymonosulfuric acid