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Characterisation of proteins from two‐dimensional electrophoresis gels by matrix‐assisted laser desorption mass spectrometry and amino acid compositional analysis
Author(s) -
Wheeler Colin H.,
Berry Sophie L.,
Wilkins Marc R.,
Corbett Joseph M.,
Ou Keli,
Gooley Andrew A.,
HumpherySmith Ian,
Williams Keith L.,
Dunn Michael J.
Publication year - 1996
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150170329
Subject(s) - peptide mass fingerprinting , mass spectrometry , chemistry , bottom up proteomics , amino acid , peptide , matrix assisted laser desorption/ionization , protein mass spectrometry , chromatography , amino acid analysis , proteome , gel electrophoresis , proteomics , two dimensional gel electrophoresis , peptide sequence , biochemistry , desorption , tandem mass spectrometry , gene , organic chemistry , adsorption
Amino acid compositional analysis and peptide mass fingerprinting by matrix assisted laser desorption mass spectrometry have been used to characterise proteins obtained from two‐dimensional electrophoresis (2‐DE) separations of human cardiac proteins. A group of twelve protein spots was selected for analysis. The identities of eight of the proteins had been determined by conventional protein characterisation methods, two were unknown proteins and two had putative identities from protein database spot comparison. Amino acid analysis and peptide mass fingerprinting gave corresponding identities for seven of the twelve proteins, which also agreed with our initial identifications. Three proteins which had been identified previously were not confirmed in this study and putative identities were obtained for the two unknown proteins. The advantages, problems and use of amino acid analysis and peptide mass fingerprinting for the analysis of proteins from 2‐DE are discussed. The data highlight the need to use orthogonal techniques for the unequivocal identification of proteins from 2‐DE gels.