Identification and characterization by high-performance liquid chromatography/electrospray ionization mass spectrometry of a new variant hemoglobin, Mataro [beta134(H12) Val > Ala.
This work illustrates the practical use of combined microbore reversed-phase high-performance liquid chromatography (RP-HPLC) with electrospray ionization mass spectrometry (ESI-MS) in protein identification. The approach consisted of the detection of the abnormal beta-globin chain by ESI-MS analysis of mixtures of intact globins, which simultaneously provided their molecular masses. Separation of the polypeptide globin chains was carried out using microbore C4 RP-HPLC on-line with ESI-MS. Direct peptide-mapping ESI-MS without previous chromatographic separation was performed in order to identify tryptic peptides from whole blood. For the sequence confirmation of the abnormal peptide containing the mutation point, C18 RP-HPLC tryptic separation of the globin mixture on-line with collision-induced dissociation (CID) fragmentation was done. The y series ions allowed the identification of the hemoglobin (Hb) variant as [beta134(H12) Val > Ala]. This new Hb was named Hb Mataró, after the city where it was detected.
Motos Gallardo
A
A
Fundació per a la Recerca Biomèdica 'Germans Trias i Pujol' (HUGTIP), Badalona, Barcelona, Spain. fgallar1@pie.xtec.es
Hernández
J A
JA
Hernández
J M
JM
Rovira
J M
JM
Fluvià
L
L
Bosch
A
A
Pastor
M C
MC
D000409:Alanine; D019943:Amino Acid Substitution; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D014644:Genetic Variation; D005914:Globins / Q000737:chemistry*; D006455:Hemoglobins, Abnormal / Q000737:chemistry*; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D010446:Peptide Fragments / Q000737:chemistry; D010449:Peptide Mapping; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D014633:Valine
D006455:Hemoglobins, Abnormal; D010446:Peptide Fragments; D005914:Globins; D014633:Valine; D000409:Alanine