βCysteine 93 in human hemoglobin: a gateway to oxidative stability in health and disease

Abstract

βcysteine 93 residue plays a key role in oxygen (O₂)-linked conformational changes in the hemoglobin (Hb) molecule. This solvent accessible residue is also a target for binding of thiol reagents that can remotely alter O₂ affinity, cooperativity, and Hb's sensitivity to changes in pH. In recent years, βCys93 was assigned a new physiological role in the transport of nitric oxide (NO) through a process of S-nitrosylation as red blood cells (RBCs) travel from lungs to tissues. βCys93 is readily and irreversibly oxidized in the presence of a mild oxidant to cysteic acid, which causes destabilization of Hb resulting in improper protein folding and the loss of heme. Under these oxidative conditions, ferryl heme (HbFe⁴⁺), a higher oxidation state of Hb is formed together with its protein radical (^.HbFe⁴⁺). This radical migrates to βCys93 and interacts with other "hotspot" amino acids that are highly susceptible to oxidative modifications. Oxidized βCys93 may therefore be used as a biomarker of oxidative stress, reflecting the deterioration of Hb within RBCs intended for transfusion or RBCs from patients with hemoglobinopathies. Site specific mutation of a redox active amino acid(s) to reduce the ferryl heme or direct chemical modifications that can shield βCys93 have been proposed to improve oxidative resistance of Hb and may offer a protective therapeutic strategy.