Abstract
Population migration has led to the global dispersion of human hemoglobinopathies and has precipitated a need for their identification. An effective mass spectrometry-based procedure involves analysis of the intact α- and β-globin chains to determine their mass, followed by location of the variant amino acid residue by direct analysis of the enzymatically digested chains and low-energy collision induced dissociation of the variant peptide. Using this procedure, a variant was identified as either β54Val→Leu or β54Val→Ile, since the amino acids leucine and isoleucine cannot be distinguished using low-energy collisions. Here, we describe how hot electron capture dissociation on a Fourier transform-ion cyclotron resonance mass spectrometer was used to distinguish isoleucine from leucine and identify the mutation as β54(D5)Val→Ile. This is a novel variant, and we have named it Hb Askew.
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Hardison, R. C.; Chui, D. H. K.; Giardine, B.; Riemer, C.; Patrinos, G. P.; Anagnou, N.; Millar, W. HbVar: A relational database of human hemoglobin variants and thalassemia mutations at the globin gene server. Human Mutat. 2002, 19, 225–233. (http://globin.cse.psu.edu).
Wild, B. J.; Green, B. N.; Cooper, E. K.; Lalloz, M. R. A.; Erten, S.; Stephens, A. D.; Layton, D. M. Rapid identification of hemoglobin variants by electrospray ionization mass spectrometry. Blood Cells Mol. Dis. 2001, 27, 691–704.
Rai, D. K.; Griffiths, W. J.; Landin, B.; Wild, B. J.; Alvelius, G.; Green, B. N. Accurate mass measurement by electrospray ionization quadrupole mass spectrometry: Detection of variants differing by <6 Da from normal in human hemoglobin heterozygotes. Anal. Chem. 2003, 75, 1978–1982.
Bateman, R. H.; Green, B. N.; Morris, M. Electrospray ionization mass spectrometric analysis of the globin chains in hemoglobin heterozygotes can detect the variants HbC, D, and E. Clin. Chem. 2008, 54, 1256–1257.
Roberts, N. B.; Green, B. N.; Morris, M. Potential of electrospray mass spectrometry for quantifying glycohemoglobin. Clin. Chem. 1997, 43, 771–778.
Roberts, N. B.; Amara, A. B.; Morris, M.; Green, B. N. Long-term evaluation of electrospray ionization mass spectrometric analysis of glycated hemoglobin. Clin. Chem. 2001, 47, 316–321.
Williams, J. P.; Jackson, H.; Green, B. N. Hb Belleville [β10(A7)Ala→Thr] affects the determination of Hb A1C by routine cation exchange high performance liquid chromatography. Hemoglobin 2009, 33, 45–50.
Williams, J. P.; Scrivens, J. H.; Green, B. N.; Farrar, L. M.; Sutcliffe, M. Hb Leeds [β56(D7)Gly→Cys]: A new hemoglobin that aggravates anemia in a child with β 0-thalassemia trait. Hemoglobin 2007, 31, 367–373.
Roepstorff, P.; Fohlman, J. Proposal for a common nomenclature for sequence ions in mass spectra of peptides. Biol. Mass Spectrom. 1984, 11, 601.
Kjeldsen, F.; Haselmann, K. F.; Budnik, B. A.; Jensen, F.; Zubarev, R. A. Dissociative capture of hot (3–13 eV electrons by polypeptide polycations: An efficient process accompanied by secondary fragmentation. Chem. Phys. Lett. 2002, 356, 201–206.
Cooper, H. J.; Hakansson, K.; Marshall, A. G. The role of electron capture dissociation in biomolecular analysis. Mass Spectrom. Rev. 2005, 24, 201–222.
Zubarev, R. A.; Kelleher, N. L.; McLafferty, F. W. ECD of multiply charged protein cations: A nonergodic process. J. Am. Chem. Soc. 1998, 120, 3265–3266.
Kjeldsen, F.; Haselmann, K. F.; Sorensen, E. S.; Zubarev, R. A. Distinguishing of Ile/Leu amino acid residues in the PP3 protein by (hot) electron capture dissociation in Fourier transform ion cyclotron resonance mass spectrometry. Anal. Chem. 2003, 75, 1267–1274.
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Published online May 18, 2009
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Williams, J.P., Creese, A.J., Roper, D.R. et al. Hot electron capture dissociation distinguishes leucine from isoleucine in a novel hemoglobin variant, Hb Askew, β54(D5)Val→Ile. J Am Soc Mass Spectrom 20, 1707–1713 (2009). https://doi.org/10.1016/j.jasms.2009.05.002
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DOI: https://doi.org/10.1016/j.jasms.2009.05.002