LTQ Orbitrap Velos in routine de novo sequencing of non‐tryptic skin peptides from the frog Rana latastei with traditional and reliable manual spectra interpretation
peer-reviewed · Rapid Communications in Mass Spectrometry · 2016
| Date | 2016-01-30 |
| Type | peer-reviewed |
| Venue | Rapid Communications in Mass Spectrometry |
| Publisher | Wiley |
| Contribution | downstream-application |
| DOI | 10.1002/rcm.7436 |
| Citations (OpenAlex) | 17 |
| Venue 2-year citedness | 1.80 |
Abstract
Rationale Mass spectrometry has shown itself to be the most efficient tool for the sequencing of peptides. However, de novo sequencing of novel natural peptides is significantly more challenging in comparison with the same procedure applied for the tryptic peptides. To reach the goal in this case it is essential to select the most efficient methods of triggering fragmentation and combine all the possible complementary techniques. Methods Collision-induced dissociation (CID), high-energy collision dissociation (HCD), and electron-transfer dissociation (ETD) tandem mass spectra recorded with a LTQ Orbitrap Velos instrument were used for the elucidation of the sequence of the natural non-tryptic peptides from the skin secretion of Rana latastei. Manual interpretation of the spectra was applied. Results The combined approach using CID, HCD, and ETD tandem mass spectra of the multiprotonated peptides in various charge states, as well as of their proteolytic fragments, allowed the sequences of seven novel peptides from the skin secretion of Rana latastei to be established. Conclusions Manual mass spectrometry sequencing of natural non-tryptic peptides from the skin secretion of Rana latastei provided the opportunity to work successfully with these species and demonstrated once again its advantage over automatic approaches.
Methods and tools
- Rana latastei skin peptidome: Routine de novo sequencing of non-tryptic frog skin peptides on an LTQ Orbitrap Velos with manual spectrum interpretation, on the argument that natural non-tryptic peptides are harder than the tryptic case automation was built for.
Cites (10)
- Lessons in de novo peptide sequencing by tandem mass spectrometry (2015) crossref
- Discrimination of Leucine and Isoleucine in Peptides Sequencing with Orbitrap Fusion Mass Spectrometer (2014) crossref
- pNovo+: De Novo Peptide Sequencing Using Complementary HCD and ETD Tandem Mass Spectra (2013) crossref
- pNovo: De novo Peptide Sequencing and Identification Using HCD Spectra (2010) crossref
- De Novo Analysis of Peptide Tandem Mass Spectra by Spectral Graph Partitioning (2006) crossref
- Proteomics-Grade de Novo Sequencing Approach (2005) crossref
- PepNovo: de novo peptide sequencing via probabilistic network modeling (2005) crossref
- Implementation and Uses of Automated de Novo Peptide Sequencing by Tandem Mass Spectrometry (2001) crossref
- De novo peptide sequencing via tandem mass spectrometry (1999) crossref
- Sequence database searches via de novo peptide sequencing by tandem mass spectrometry (1997) crossref