Y. Wada et al., Tandem mass spectrometric analysis of C-13-containing ions from a mixture of homologous peptides differing by one mass unit at a residue, J MASS SPEC, 35(2), 2000, pp. 242-250
Tandem mass spectrometry of a mixture of two peptides that differ from each
other by a single mass unit due to mutation is presented. The mutant beta-
globin of hemoglobin Hoshida is present along with the normal counterpart,
and the amino acid substitution of glutamine for glutamic acid is located w
ithin tryptic peptide T5 of M-r 2057.9. The mass of the mutated peptide is
1 u lower. In the isotopic cluster for the doubly charged ion of the peptid
e T5, the resolved ion with mass of 1030.0 represents the normal peptide wi
th 93 C-12 atoms and the mutated one with 92 C-12 and one C-13 atoms. Colli
sion-induced dissociation (CID) of this composite ion identified the mutati
on by presenting a key fragment derived from the C-12-only mutant peptide,
as reported in a previous study. Similarly, when an ion containing multiple
C-13 atoms was selected as a precursor for CID, the mutation could be iden
tified, even in large fragments, by a marked change in the shape of the iso
topic cluster for the consecutive product ions. This study demonstrates the
merit of selecting a resolved ion rather than the whole isotopic cluster a
s a precursor in the CID measurements of large peptides or proteins for cha
racterizing heterozygous mutations. Copyright (C) 2000 John Wiley & Sons, L
td.