Combined mass spectrometric methods for the characterization of human hemoglobin variants localized within alpha T9 peptide: identification of Hb Villeurbanne alpha 89 (FG1) His-->Tyr.
Mutation-induced amino acid exchanges occurring on the large T9 peptide of the alpha-chain of human hemoglobin (residues 62-90) are difficult to identify. Despite their high m/z value (around m/z 3000), collision-induced dissociation spectra of liquid secondary ion mass spectrometrically generated protonated alpha T9 peptides were performed successfully. In parallel electrospray mass spectrometry (MS) was used both to measure the molecular mass of the intact proteins and to determine the number of protonatable sites in the alpha T9 peptides. Peptide ladder sequencing using carboxypeptidase digestions and analysis of the truncated peptides by matrix-assisted laser desorption ionization time-of-flight MS confirmed the interpretation. This set of methods allowed the characterization of three hemoglobin variants, with amino acid exchanges located in the alpha T9 part of the sequence. Two of them, Hb Aztec [alpha 76(EF5) Met-->Thr] and Hb M-Iwate [alpha 87(F8) His-->Tyr] were already known. The third [alpha 89(FG1) His-->Tyr] was novel and named Hb Villeurbanne.
Déon
C
C
Institut de Pharmacologie et Biologie Structurale du CNRS, Toulouse, France.
Promé
J C
JC
Promé
D
D
Francina
A
A
Groff
P
P
Kalmes
G
G
Galactéros
F
F
Wajcman
H
H
D000595:Amino Acid Sequence; D002268:Carboxypeptidases / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004588:Electrophoresis, Cellulose Acetate; D005914:Globins / Q000737:chemistry / Q000235:genetics; D006449:Hemoglobin M / Q000737:chemistry / Q000235:genetics; D006455:Hemoglobins, Abnormal / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009154:Mutation; D010455:Peptides / Q000737:chemistry; D017421:Sequence Analysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000378:metabolism
D006455:Hemoglobins, Abnormal; D010455:Peptides; C107881:hemoglobin Villeurbanne; C049100:hemoglobin Aztec; D005914:Globins; C027542:hemoglobin M Iwate; D006449:Hemoglobin M; D002268:Carboxypeptidases; D014357:Trypsin