Journal
EXPERT REVIEW OF PROTEOMICS
Volume 3, Issue 6, Pages 585-596Publisher
TAYLOR & FRANCIS LTD
DOI: 10.1586/14789450.3.6.585
Keywords
CAD; collision-activated dissociation; ECD; electron-capture dissociation; electrospray-ionization; intact mass tags; intact proteins; integral membrane proteins; mass spectrometry; post-transcriptional modifications; post-translational modifications; top-down proteomics
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Top-down mass spectrometry focuses on intact proteins, thereby avoiding loss of information accompanying 'shotgun' protocols that reduce the proteome to a collection of peptides. A suite of liquid-chromatography technologies has been developed for purification of intact integral membrane proteins in aqueous/organic solvent mixtures compatible with biological 'soft-ionization' mass spectrometry, preserving covalent structure into the gas phase, Multiply charged protein ions are fragmented in the gas phase, using either collision-activated or electron-capture dissociation, thus yielding complex spectra of sequence-dependent product ions that collectively define the original native covalent state of an intact protein. Top down offers a more detail-orientated approach to post-transcriptional and post-translational diversity allowing an enhanced insight beyond genomic translation, which has now extended into the bilayer proteome.
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