Simplifying Fragmentation Patterns of Multiply Charged Peptides by N-Terminal Derivatization and Electron Transfer Collision Activated Dissociation
peer-reviewed · Analytical Chemistry · 2009
| Date | 2009-05-01 |
| Type | peer-reviewed |
| Venue | Analytical Chemistry |
| Publisher | American Chemical Society (ACS) |
| Contribution | adjacent |
| DOI | 10.1021/ac9000942 |
| Citations (OpenAlex) | 45 |
| Venue 2-year citedness | 7.10 |
Abstract
N-terminal peptide derivatization strategies used in conjunction with tandem mass spectrometry to yield simplified fragmentation patterns have shown limited success for the de novo sequencing of multiply charged peptides, including those predominantly formed in LC-ESI-MS experiments. Significant proton mobilization occurs for multiply charged peptides upon collisional activation, resulting in the formation of both N-terminal and C-terminal product ions rather than an exclusive series of C-terminal ions preferred for de novo sequencing algorithms. To circumvent this problem, multiply charged, N-terminally derivatized peptides were subjected to electron transfer reactions with fluoranthene anions to produce singly charged, radical species. Upon subsequent “soft” collision induced dissociation (CID), highly abundant z-type ions were formed nearly exclusively, which yielded simplified fragmentation patterns amenable to de novo sequencing methods. Furthermore, the derivatized peptides retained labile phosphoric acid moieties, and the enhanced set of z ions were also observed for peptides not possessing basic C-terminal residues, a type of peptide that poses more challenges to traditional simplification methods based on collision activated dissociation. This improved LC-MS(n) strategy was demonstrated for a variety of multiply charged model peptides and a tryptic digest of myoglobin.
Methods and tools
- Derivatization plus ETcaD de novo: N-terminally derivatized, multiply charged peptides are charge-reduced by electron transfer and then collisionally activated to give almost only z-ions for de novo sequencing.
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Cited by (7)
- UVnovo: A de Novo Sequencing Algorithm Using Single Series of Fragment Ions via Chromophore Tagging and 351 nm Ultraviolet Photodissociation Mass Spectrometry (2016) both
- Bromine isotopic signature facilitates de novo sequencing of peptides in free‐radical‐initiated peptide sequencing (FRIPS) mass spectrometry (2015) crossref
- Neutron-encoded Signatures Enable Product Ion Annotation From Tandem Mass Spectra (2013) both
- De Novo Sequencing of Peptides Using Selective 351 nm Ultraviolet Photodissociation Mass Spectrometry (2013) crossref
- High-Confidence de Novo Peptide Sequencing Using Positive Charge Derivatization and Tandem MS Spectra Merging (2013) crossref
- 193 nm Ultraviolet Photodissociation of Imidazolinylated Lys-N Peptides for De Novo Sequencing (2012) crossref
- Dimethyl isotope labeling assisted de novo peptide sequencing (2010) crossref