Establishment of a combined strategy of genetic and mass spectrometric analyses for characterizing hemoglobin mutations. An example of Hb Hoshida (beta43Glu-->Gln).
Structural analysis of mutant hemoglobins has been efficiently accomplished by a consecutive mass spectrometric strategy: molecular mass measurement of the protein to detect mutation and to determine the molecular mass change, and fragmentation analysis with collision-induced dissociation to determine the site and type of mutation. A flaw of this method is an inherent inability to detect a mutation associated with no or little change of the molecular mass. In the present study, the strategy was improved by incorporating genetic analysis prior to mass spectrometry, which confirms the resulting amino acid change and searches for possible post-translational modifications. The method was applied to an unknown mutant, and elucidated a substitution of glutamine for glutamic acid at position 43 of beta-globin subunit, the mutation of Hb Hoshida.
Kaneko
R
R
Department of Physics, Graduate School of Science, Osaka University, Toyonaka, Japan.
Wada
Y
Y
Hisada
M
M
Naoki
H
H
Matsuo
T
T
D000595:Amino Acid Sequence; D001483:Base Sequence; D017931:DNA Primers; D005260:Female; D005820:Genetic Testing / Q000379:methods*; D018698:Glutamic Acid / Q000235:genetics*; D005998:Glycine / Q000235:genetics*; D006455:Hemoglobins, Abnormal / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D007525:Isoelectric Focusing; D013058:Mass Spectrometry / Q000379:methods*; D008875:Middle Aged; D008969:Molecular Sequence Data; D017354:Point Mutation
D017931:DNA Primers; D006455:Hemoglobins, Abnormal; D018698:Glutamic Acid; C074595:hemoglobin Hoshida; D005998:Glycine