Open AccessAlgorithm of Protein Sequence Determination by Combination of the Edman Degradation Method and Shotgun Mass Spectrometry
Author(s) -
V. S. Skvortsov,
A.V. Mikurova,
Nikita E. Vavilov,
В. Г. Згода
Publication year - 2018
Publication title -
biomedical chemistry: research and methods
Language(s) - English
Resource type - Journals
ISSN - 2618-7531
DOI - 10.18097/bmcrm00087
Subject(s) - edman degradation , shotgun proteomics , mass spectrometry , chromatography , trypsin , chemistry , shotgun , protein sequencing , peptide sequence , protein mass spectrometry , sequence (biology) , tandem mass spectrometry , proteomics , biochemistry , enzyme , gene
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.