Identification and characterization by high-performance liquid chromatography/electrospray ionization mass spectrometry of a new variant hemoglobin, Mataro [beta 134(H12) Val > Ala]

This work illustrates the practical use of combined microbore reversed-phas e high-performance liquid chromatography (RP-HPLC) with electrospray ioniza tion mass spectrometry (ESI-MS) in protein identification. The approach con sisted of the detection of the abnormal beta -globin chain by ESI-MS analys is of mixtures of intact globins, which simultaneously provided their molec ular masses. Separation of the polypeptide globin chains was carried out us ing microbore C-4 RP-HPLC on-line with ESI-MS. Direct peptide-mapping ESI-M S without previous chromatographic separation was performed in order to ide ntify tryptic peptides from whole blood. For the sequence confirmation of t he abnormal peptide containing the mutation point, C-18 RP-HPLC tryptic sep aration of the globin mixture on-line with collision-induced dissociation ( CID) fragmentation was done. The y series ions allowed the identification o f the hemoglobin (Hb) variant as [beta 134(H12) Val > Ala]. This new Hb was named Rb Matar6, after the city where it was detected. Copyright (C) 2001 John Wiley & Sons, Ltd.