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Updated: Aug 4, 2026

Measurement of Heme Synthesis Levels in Mammalian Cells
Published on: July 9, 2015
Identification of a nickel(II) binding site on hemoglobin which confers susceptibility to oxidative deamination and
J Levine1, M Weickert, M Pagratis
1Somatogen Inc., Boulder, Colorado 80301, USA.
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.
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