Dimethyl isotope labeling assisted de novo peptide sequencing
peer-reviewed · Journal of the American Society for Mass Spectrometry · 2010
| Date | 2010-12-01 |
| Type | peer-reviewed |
| Venue | Journal of the American Society for Mass Spectrometry |
| Publisher | American Chemical Society (ACS) |
| Contribution | adjacent |
| DOI | 10.1016/j.jasms.2010.08.007 |
| Citations (OpenAlex) | 27 |
| Venue 2-year citedness | 2.58 |
Abstract
Here, we explore a de novo sequencing strategy in which we combine Lys-N protein digestion with differential isotopic dimethyl labeling to facilitate the (de novo) identification of multiply charged peptides in ESI-MS, both under CID and ETD conditions. For a large fraction of the Lys-N generated peptides, all primary amines are present at the N-terminal lysine, enabling specific labeling of the N-terminus. Differential derivatization of only the peptide N-terminus in combination with the simultaneous fragmentation of the corresponding isotopologues allows the straightforward distinction of N-terminal fragments from C-terminal and internal fragments. Furthermore, also singly and multiply charged N-terminal fragments can easily be distinguished due to the mass differences of the isotope labeled fragment pairs. As a proof of concept, we applied this approach to proteins isolated from an avocado fruit, and were able to partially de novo sequence and correctly align, with green plant homologues, a previously uncharacterized avocado ascorbate peroxidase.
Methods and tools
- Dimethyl isotope labeling de novo sequencing: Dimethyl isotope labeling-assisted workflow for de novo peptide sequencing.
Cites (11)
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