Bromine isotopic signature facilitates de novo sequencing of peptides in free‐radical‐initiated peptide sequencing (FRIPS) mass spectrometry
peer-reviewed · Journal of Mass Spectrometry · 2015
| Date | 2015-02-01 |
| Type | peer-reviewed |
| Venue | Journal of Mass Spectrometry |
| Publisher | Wiley |
| Contribution | adjacent |
| DOI | 10.1002/jms.3539 |
| Citations (OpenAlex) | 8 |
| Venue 2-year citedness | 1.85 |
Abstract
We recently showed that free-radical-initiated peptide sequencing mass spectrometry (FRIPS MS) assisted by the remarkable thermochemical stability of (2,2,6,6-tetramethyl-piperidin-1-yl)oxyl (TEMPO) is another attractive radical-driven peptide fragmentation MS tool. Facile homolytic cleavage of the bond between the benzylic carbon and the oxygen of the TEMPO moiety in o-TEMPO-Bz-C(O)-peptide and the high reactivity of the benzylic radical species generated in •Bz-C(O)-peptide are key elements leading to extensive radical-driven peptide backbone fragmentation. In the present study, we demonstrate that the incorporation of bromine into the benzene ring, i.e. o-TEMPO-Bz(Br)-C(O)-peptide, allows unambiguous distinction of the N-terminal peptide fragments from the C-terminal fragments through the unique bromine doublet isotopic signature. Furthermore, bromine substitution does not alter the overall radical-driven peptide backbone dissociation pathways of o-TEMPO-Bz-C(O)-peptide. From a practical perspective, the presence of the bromine isotopic signature in the N-terminal peptide fragments in TEMPO-assisted FRIPS MS represents a useful and cost-effective opportunity for de novo peptide sequencing.
Methods and tools
- Br-TEMPO FRIPS de novo sequencing: Adds bromine to the free-radical-initiated peptide sequencing reagent so N-terminal fragments carry a doublet signature that eases de novo reading.
Cites (18)
- C-terminal de novo sequencing of peptides using oxazolone-based derivatization with bromine signature (2011) crossref
- Dimethyl isotope labeling assisted de novo peptide sequencing (2010) crossref
- pNovo: De novo Peptide Sequencing and Identification Using HCD Spectra (2010) crossref
- De novo sequencing of peptides by MS/MS (2010) crossref
- De novo peptide sequencing by tandem MS using complementary CID and electron transfer dissociation (2009) crossref
- Simplifying Fragmentation Patterns of Multiply Charged Peptides by N-Terminal Derivatization and Electron Transfer Collision Activated Dissociation (2009) crossref
- Straightforward and de Novo Peptide Sequencing by MALDI-MS/MS Using a Lys-N Metalloendopeptidase (2009) crossref
- Specific isolation of N‐terminal fragments from proteins and their high‐fidelity de novo sequencing (2007) crossref
- De Novo Peptide Sequencing and Identification with Precision Mass Spectrometry (2007) crossref
- N-Terminal amino acid side-chain cleavage of chemically modified peptides in the gas phase: A mass spectrometry technique for N-terminus identification (2006) crossref
- Peptide de Novo Sequencing Facilitated by a Dual-Labeling Strategy (2005) crossref
- Highly informative proteome analysis by combining improved N ‐terminal sulfonation for de novo peptide sequencing and online capillary reverse‐phase liquid chromatography/tandem mass spectrometry (2004) crossref
- De novo sequencing, peptide composition analysis, and composition-based sequencing: A new strategy employing accurate mass determination by Fourier transform ion cyclotron resonance mass spectrometry (2004) crossref
- Peptide and protein de novo sequencing by mass spectrometry (2003) crossref
- Use of Deuterium-Labeled Lysine for Efficient Protein Identification and Peptide de Novo Sequencing (2002) crossref
- Automatedde novo sequencing of proteins using the differential scanning technique (2001) crossref
- Quantitation and Facilitated de Novo Sequencing of Proteins by Isotopic N-Terminal Labeling of Peptides with a Fragmentation-Directing Moiety (2000) crossref
- A method for high-sensitivity peptide sequencing using postsource decay matrix-assisted laser desorption ionization mass spectrometry (1999) crossref