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Use of 18O Labels to monitor deamidation during protein and peptide sample processing
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Li, Xiaojuan, Cournoyer, Jason J., Lin, Cheng and O'Connor, Peter B. (2008) Use of 18O Labels to monitor deamidation during protein and peptide sample processing. Journal of The American Society for Mass Spectrometry, Vol.19 (No.6). pp. 855-864. doi:10.1016/j.jasms.2008.02.011 ISSN 1044-0305.
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Official URL: http://dx.doi.org/10.1016/j.jasms.2008.02.011
Abstract
Nonenzymatic deamidation of asparagine residues in proteins generates aspartyl (Asp) and isoaspartyl (isoAsp) residues via a succinimide intermediate in a neutral or basic environment. Electron capture dissociation (ECD) can differentiate and quantify the relative abundance of these isomeric products in the deamidated proteins. This method requires the proteins to be digested, usually by trypsin, into peptides that are amenable to ECD. ECD of these peptides can produce diagnostic ions for each isomer; the c · + 58 and z − 57 fragment ions for the isoAsp residue and the fragment ion ((M + nH)(n−1)+· − 60) corresponding to the side-chain loss from the Asp residue. However, deamidation can also occur as an artifact during sample preparation, particularly when using typical tryptic digestion protocols. With 18O labeling, it is possible to differentiate deamidation occurring during trypsin digestion which causes a +3 Da (18O1 + 1D) mass shift from the pre-existing deamidation, which leads to a +1-Da mass shift. This paper demonstrates the use of 18O labeling to monitor three rapidly deamidating peptides released from proteins (calmodulin, ribonuclease A, and lysozyme) during the time course of trypsin digestion processes, and shows that the fast (∼4 h) trypsin digestion process generates no additional detectable peptide deamidations.
| Item Type: | Journal Article | ||||
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| Subjects: | Q Science > QD Chemistry Q Science > QP Physiology |
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| Divisions: | Faculty of Science, Engineering and Medicine > Science > Chemistry | ||||
| Library of Congress Subject Headings (LCSH): | Deamination, Tandem mass spectrometry, Proteins -- Deamination, Peptides -- Analysis, Proteomics | ||||
| Journal or Publication Title: | Journal of The American Society for Mass Spectrometry | ||||
| Publisher: | Springer New York LLC | ||||
| ISSN: | 1044-0305 | ||||
| Official Date: | 2008 | ||||
| Dates: |
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| Volume: | Vol.19 | ||||
| Number: | No.6 | ||||
| Number of Pages: | 10 | ||||
| Page Range: | pp. 855-864 | ||||
| DOI: | 10.1016/j.jasms.2008.02.011 | ||||
| Status: | Peer Reviewed | ||||
| Publication Status: | Published | ||||
| Access rights to Published version: | Open Access (Creative Commons) | ||||
| Funder: | National Institutes of Health (U.S.) (NIH), National Institute of General Medical Sciences (U.S.) (NIGMS), National Heart, Lung, and Blood Institute (NHLBI), National Center for Research Resources (U.S.) (NCRR), MDS Sciex, Petroleum Research Fund | ||||
| Grant number: | P41RR10888 (NIH/NCRR), N01HV28178 (NIH/NHLBI), R01GM078293 (NIH/NIGMS) |
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