A method for N-terminal de novo sequence analysis of proteins by matrix-assisted laser desorption/ionization mass spectrometry
peer-reviewed · Analytical Biochemistry · 2008
| Date | 2008-09-01 |
| Type | peer-reviewed |
| Venue | Analytical Biochemistry |
| Publisher | Elsevier BV |
| Contribution | algorithm |
| DOI | 10.1016/j.ab.2008.05.053 |
| Citations (OpenAlex) | 35 |
Abstract
A novel method for isolation and de novo sequencing of N-terminal peptides from proteins is described. The method presented here combines selective chemical tagging using succinimidyloxycarbonylmethyl tris(2,4,6-trimethoxyphenyl)phosphonium bromide (TMPP-Ac-OSu) at the N(alpha)-amino group of peptides after digestion by metalloendopeptidase (from Grifola frondosa) and selective capture procedures using p-phenylenediisothiocyanate resin, by which the N-terminal peptide can be isolated, whether or not it is N-terminally blocked. The isolated N-terminal peptide modified N-terminally with TMPP-Ac-OSu reagent produces a simple fragmentation pattern under tandem mass spectrometric analysis to significantly facilitate sequencing.
Methods and tools
- TMPP N-terminal peptide de novo sequencing: Tags peptide N-termini with TMPP-Ac-OSu after metalloendopeptidase digestion and isolates them, giving MALDI spectra of N-terminal peptides clean enough to sequence de novo.