Hart, Sarah R. and Wai Lau, King Wai and Hao, Zhiqi and Broadhead, Richard and Portman, Neil and Huhmer, Andreas and Gull, Keith and McKean, Paul G. and Hubbard, Simon J. and Gaskell, Simon J. (2009) Analysis of the trypanosome flagellar proteome using a combined electron transfer collisionally activated dissociation strategy. Journal of the American Society for Mass Spectrometry, 20 (2). pp. 167-175. ISSN 1044-0305Full text not available from this repository.
The use of electron-transfer dissociation as an alternative peptide ion activation method for generation of protein sequence information is examined here in comparison with the conventional method of choice, collisionally activated dissociation, using a linear ion trapping instrument. Direct comparability between collisionally and electron-transfer-activated product ion data were ensured by employing an activation-switching method during acquisition, sequentially activating precisely the same precursor ion species with each fragmentation method in turn. Sequest (Thermo Fisher Scientific, San Jose, CA) searching of product ion data generated an overlapping yet distinct pool of polypeptide identifications from the products of collisional and electron-transfer-mediated activation products. To provide a highly confident set of protein recognitions, identification data were filtered using parameters that achieved a peptide false discovery rate of 1%, with two or more independent peptide assignments required for each protein. The use of electron transfer dissociation (ETD) has allowed us to identify additional peptides where the quality of product ion data generated by collisionally activated dissociation (CAD) was insufficient to infer peptide sequence. Thus, a combined ETD/CAD approach leads to the recognition of more peptides and proteins than are achieved using peptide analysis by CAD- or ETD-based tandem mass spectrometry alone.
|Journal or Publication Title:||Journal of the American Society for Mass Spectrometry|
|Subjects:||Q Science > QH Natural history > QH301 Biology|
|Departments:||Faculty of Science and Technology > Lancaster Environment Centre|
Faculty of Health and Medicine > Biomedical & Life Sciences
|Deposited By:||Dr Paul G McKean|
|Deposited On:||12 Mar 2009 09:21|
|Last Modified:||26 Jul 2012 16:28|
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