Abstract
An electrospray mass spectrometric approach to the identification of a human hemoglobin (Hb) variant involving a Cys residue incorporation is presented. In Hb Ta-Li (β83Gly → Cys), Cys83 forms inter-molecular disulfide bridges. Routine analysis of the denatured Hb showed the presence of a minor β chain variant whose mass apparently was 1 Da less than the expected mass difference of 46 Da for a Gly → Cys substitution. Reduction of the globin chains with dithiothreitol gave an intense monomer with the expected mass difference for the Gly → Cys substitution. After reprocessing the original raw data from the denatured Hb and taking into account the possibility of dimer formation, a component was revealed whose mass was consistent with a disulfide linked dimer of Ta-Li β globins. The mutation was localized to peptide βT10 by analysis of a tryptic digest. Tandem mass spectrometry and DNA sequencing confirmed the Gly → Cys substitution occurred at residue 83 of the β chain. Problems encountered in identifying the components in mixtures of monomers and dimers are discussed.
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Rai, D.K., Landin, B., Griffiths, W.J. et al. Characterization of the elusive disulfide bridge forming human Hb variant: Hb Ta-Li β83 (EF7)Gly → Cys by electrospray mass spectrometry. J Am Soc Mass Spectrom 13, 187–191 (2002). https://doi.org/10.1016/S1044-0305(01)00349-X
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DOI: https://doi.org/10.1016/S1044-0305(01)00349-X