Journal
JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY
Volume 32, Issue 7, Pages 1752-1758Publisher
AMER CHEMICAL SOC
DOI: 10.1021/jasms.1c00113
Keywords
top-down mass spectrometry; electron capture dissociation; electron ionization dissociation; UVPD; internal fragments; CAD
Funding
- US National Institutes of Health [R01GM103479, S10RR028893]
- US National Science Foundation (NSF) [CHE1808492]
- US Department of Energy [DE-FC02-02ER63421]
- Ruth L. Kirschstein National Research Service Award program [GM007185]
- NIH [P41 GM103533]
- NSF [CHE1904577]
Ask authors/readers for more resources
Analysis of internal fragments in TD-MS experiments can complement terminal fragments, increase protein sequence coverage, and enable more reliable localization of modification sites.
Top-down mass spectrometry (TD-MS) of intact proteins results in fragment ions that can be correlated to the protein primary sequence. Fragments generated can either be terminal fragments that contain the N- or C-terminus or internal fragments that contain neither termini. Traditionally in TD-MS experiments, the generation of internal fragments has been avoided because of ambiguity in assigning these fragments. Here, we demonstrate that in TD-MS experiments internal fragments can be formed and assigned in collision-based, electron-based, and photonbased fragmentation methods and are rich with sequence information, allowing for a greater extent of the primary protein sequence to be explained. For the three test proteins cytochrome c, myoglobin, and carbonic anhydrase II, the inclusion of internal fragments in the analysis resulted in approximately 15-20% more sequence coverage, with no less than 85% sequence coverage obtained. Combining terminal fragment and internal fragment assignments results in near complete protein sequence coverage. Hence, by including both terminal and internal fragment assignments in TD-MS analysis, deep protein sequence analysis, allowing for the localization of modification sites more reliably, can be possible.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available