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Mass spectrometic detection for capillary isoelectric focusing separations of complex protein mixtures
Author(s) -
Jensen Pamela K.,
PaìaTolić Ljiljana,
Peden Kim K.,
Martinović Suzana,
Lipton Mary S.,
Anderson Gordon A.,
Tolić Nikola,
Wong KwongKwok,
Smith Richard D.
Publication year - 2000
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/(sici)1522-2683(20000401)21:7<1372::aid-elps1372>3.0.co;2-y
Subject(s) - isoelectric focusing , chromatography , capillary action , chemistry , capillary electrophoresis , isoelectric point , analytical chemistry (journal) , materials science , biochemistry , enzyme , composite material
Capillary isoelectric focusing (CIEF) can provide high‐resolution separations of complex protein mixtures, but until recently it has primarily been used with conventional UV detection. This technique would be greatly enhanced by much more information‐rich detection methods that can aid in protein characterization. We describe progress in the development of the combination of CIEF with Fourier transform ion cyclotron resonance (FTICR) mass spectrometry and its application to proteome characterization. Studies have revealed 400—1000 putative proteins in the mass range of 2—100 kDa from total injections of ˜ 300 ng protein in single CIEF‐FTICR analyses of cell lysates for both Escherichia coli (E. coli) and Deinococcus radiodurans (D. radiodurans) . We also demonstrate the use of isotope labeling of the cell growth media to improve mass measurement accuracy and provide a means for quantitative proteome‐wide measurements of protein expression. The ability to make such comprehensive and precise measurements of differences in protein expression in response to cellular perturbations should provide new insights into complex cellular processes.