Optimization of carbamylation conditions and study on the effects on the product ions of carbamylation and dual modification of the peptide by Q-TOF MS
peer-reviewed · European Journal of Mass Spectrometry · 2018
| Date | 2018-10-01 |
| Type | peer-reviewed |
| Venue | European Journal of Mass Spectrometry |
| Publisher | SAGE Publications |
| Contribution | adjacent |
| DOI | 10.1177/1469066718788665 |
| Citations (OpenAlex) | 1 |
Abstract
Modified peptides fragmented by collision-induced dissociation can offer additional sequence information, which is beneficial for the de novo sequencing of peptides. Here, the model peptide VQGESNDLK was carbamylated. The optimal conditions were as follows: temperature of 90℃, pH of 7, and the time of 60 min. Then, we studied the b- and y-series ions of the native, carbamylated, and dual-modified peptides. The results were as follows. The short carbamylated peptides (≤10 amino acid residues) produced more b-series ions (including b 1 ion). The long carbamylated peptides (>10 amino acid residues) produced additional b 1 ion but fewer y-series ions (especially in the high-mass region). The short dual-modified peptides produced more b-series ions (including b 1 ion) and more y-series ions, and their peptide sequence coverage was almost 100%. The long dual-modified peptides produce b 1 ion and more y-series ions, and their peptide sequence coverage was nearly above 90%. Therefore, both carbamylation and the dual modification method could be used to identify the N-terminal amino acid, and the dual modification method was also excellent for the de novo sequencing of the tryptic peptides.
Methods and tools
- Carbamylation-assisted de novo sequencing: Studies carbamylation and dual modification of peptides to enrich b- and y-ion series, including b1, for easier de novo sequencing by Q-TOF.