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Fast atom bombardment mass spectrometric analysis of haemoglobin variants: Use of V‐8 protease in the identification of Hb M Hyde Park and Hb San Jose
Author(s) -
Pucci P.,
Ferranti P.,
Malorni A.,
Marino G.
Publication year - 1990
Publication title -
biomedical and environmental mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 0887-6134
DOI - 10.1002/bms.1200190908
Subject(s) - protease , fast atom bombardment , chemistry , proteolytic enzymes , mass spectrometry , peptide , enzyme , trypsin , biochemistry , chromatography
The characterization of two human haemoglobin variants, Hb M Hyde Park and Hb San José, by fast atom bombardment mass spectrometry is reported. The identification of the site and nature of the amino acid substitution was performed by analysis of the peptide mixture generated by proteolytic digestion of the variant β‐globin chains with V‐8 protease. The use of this protease was instrumental in the unambiguous identification of the replaced residues because the tryptic map alone was unable to unequivocally locate the modification. Spectra obtained were easily interpreted and the characterization of Hb M Hyde Park (β, 92 Hys → Tyr) and Hb San José (β 7 Glu → Gly) was accomplished essentially by the same procedure already described for the tryptic map. These results are suggestive of alternative approaches in haemoglobin variants characterization using different proteolytic enzymes when tryptic data alone do not lead to unambiguous results.