Identification and characterization by high-performance liquid chromatography/electrospray ionization mass spectrometry of a new variant hemoglobin, Mataro [β134(H12) Val > Ala]

✍ Scribed by A. Motos Gallardo; J. A. Hernández; J. M. Hernández; J. M. Rovira; L. Fluvià; A. Bosch; M. C. Pastor

• Publisher: John Wiley and Sons
• Year: 2001
• Tongue: English
• Weight: 171 KB
• Volume: 36
• Category: Article
• ISSN: 1076-5174
• DOI: 10.1002/jms.198

✦ Synopsis

Abstract

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 β‐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 C~4~ 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, C~18~ 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 [β134(H12) Val > Ala]. This new Hb was named Hb Mataró, after the city where it was detected. Copyright © 2001 John Wiley & Sons, Ltd.