Algorithm of Protein Sequence Determination by Combination of the Edman Degradation Method and Shotgun Mass Spectrometry
peer-reviewed · Biomedical Chemistry: Research and Methods · 2018
| Date | 2018-12-01 |
| Type | peer-reviewed |
| Venue | Biomedical Chemistry: Research and Methods |
| Publisher | Institute of Biochemistry |
| Contribution | adjacent |
| DOI | 10.18097/bmcrm00087 |
| Citations (OpenAlex) | 0 |
| Venue 2-year citedness | 0.48 |
Abstract
An algorithm combining advantages of the Edman degradation method and de novo mass-spectrometric sequencing was developed. The protein from the “Diaskintest” diagnostic test was used for analysis. The protein was digested with trypsin and 5 steps of Edman degradation were carried out sequentially for the mixture of peptides. At each stage, the resulting mixture was analyzed by shotgun mass spectrometry analysis. The results of mass-spectrometry were analyzed both by the well-known de novo sequencing programs Novor and PepNovo+, and by own program that clustered individual spectra with a C-terminal signature formed by Y-ions. This approach allows us to determine confidently the amino acid sequence of the N-terminal part of the peptides obtained after the protein hydrolysis by trypsin.
Methods and tools
- Edman-assisted shotgun de novo sequencing: Runs successive Edman steps on a tryptic digest and de novo sequences each stage by MS to fix the N-terminal residues of peptides.