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Top‐down protein characterization facilitated by ion/ion reactions on a quadrupole/time of flight platform
Author(s) -
Huang Tengyi,
McLuckey Scott A.
Publication year - 2010
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201000187
Subject(s) - time of flight , quadrupole time of flight , ion , characterization (materials science) , quadrupole , chemistry , nanotechnology , materials science , chromatography , physics , atomic physics , electrospray ionization , organic chemistry
In comparison to bottom‐up proteomics approaches, whereby peptides derived from proteolytic digestion are analyzed, top‐down approaches, involving direct analysis of intact proteins, provide higher specificity for protein identification and are better‐suited for the characterization of sequence variants. However, top‐down protein characterization usually requires more sophisticated instrumentation and methodologies to deal with the more complex tandem mass spectra derived from dissociation of high mass multiply charged intact proteins. Gas‐phase ion/ion reactions are universally applicable and have proved to be useful in mixture analysis and top‐down biomolecule characterization. The coupling of the ion/ion proton transfer reaction in the context of MS/MS has been demonstrated to expand informing power in top‐down protein characterization, particularly with platforms that employ electrodynamic ion trap and TOF mass analysis. In addition, probing protein primary structure using ion/ion electron transfer dissociation usually provides extensive structurally informative fragmentation and also allows for the localization of labile PTMs. Here, the performance of the widely used quadrupole/TOF platform, equipped with ion/ion reaction functionality, for top‐down protein characterization is summarized, and various methodologies employing ion/ion reactions are reviewed.