Identification of hemoglobin variants by top-down mass spectrometry using selected diagnostic product ions.
Hemoglobin disorder diagnosis is a complex procedure combining several analytical steps. Due to the lack of specificity of the currently used protein analysis methods, the identification of uncommon hemoglobin variants (proteoforms) can become a hard task to accomplish. The aim of this work was to develop a mass spectrometry-based approach to quickly identify mutated protein sequences within globin chain variants. To reach this goal, a top-down electron transfer dissociation mass spectrometry method was developed for hemoglobin β chain analysis. A diagnostic product ion list was established with a color code strategy allowing to quickly and specifically localize a mutation in the hemoglobin β chain sequence. The method was applied to the analysis of rare hemoglobin β chain variants and an (A)γ-β fusion protein. The results showed that the developed data analysis process allows fast and reliable interpretation of top-down electron transfer dissociation mass spectra by nonexpert users in the clinical area.
Department of Human Protein Sciences, Faculty of Medicine, Geneva University, Rue Michel Servet 1, 1211, Geneva, Switzerland.
Didia
D
Coelho Graça
Ralf
R
Hartmer
Wolfgang
W
Jabs
Photis
P
Beris
Lorella
L
Clerici
Carsten
C
Stoermer
Kaveh
K
Samii
Denis
D
Hochstrasser
Yury O
YO
Tsybin
Alexander
A
Scherl
Pierre
P
Lescuyer
D000595:Amino Acid Sequence; D005319:Fetal Hemoglobin / Q000032:analysis / Q000235:genetics; D050939:Gene Fusion; D014644:Genetic Variation; D006454:Hemoglobins / Q000032:analysis* / Q000235:genetics; D006455:Hemoglobins, Abnormal / Q000032:analysis / Q000235:genetics; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D009154:Mutation; D053719:Tandem Mass Spectrometry / Q000379:methods; D057188:Workflow; D055544:beta-Globins / Q000032:analysis / Q000235:genetics
D006454:Hemoglobins; D006455:Hemoglobins, Abnormal; D055544:beta-Globins; C040843:hemoglobin Kenya; D005319:Fetal Hemoglobin