De novo sequencing of proteins by mass spectrometry

peer-reviewed · Expert Review of Proteomics · 2020

peer-reviewed · Expert Review of Proteomics · 2020. Rui Vitorino et al. Introduction Proteins are crucial for every cellular activity and unraveling their sequence and structure is…
Date 2020-10-01
Type peer-reviewed
Venue Expert Review of Proteomics
Publisher Taylor & Francis
Contribution review
DOI 10.1080/14789450.2020.1831387
Citations (OpenAlex) 42
Venue 2-year citedness 3.10

Abstract

Introduction Proteins are crucial for every cellular activity and unraveling their sequence and structure is a crucial step to fully understand their biology. Early methods of protein sequencing were mainly based on the use of enzymatic or chemical degradation of peptide chains. With the completion of the human genome project and with the expansion of the information available for each protein, various databases containing this sequence information were formed.Areas covered De novo protein sequencing, shotgun proteomics and other mass-spectrometric techniques, along with the various software are currently available for proteogenomic analysis. Emphasis is placed on the methods for de novo sequencing, together with potential and shortcomings using databases for interpretation of protein sequence data.Expert opinion As mass-spectrometry sequencing performance is improving with better software and hardware optimizations, combined with user-friendly interfaces, de-novo protein sequencing becomes imperative in shotgun proteomic studies. Issues regarding unknown or mutated peptide sequences, as well as, unexpected post-translational modifications (PTMs) and their identification through false discovery rate searches using the target/decoy strategy need to be addressed. Ideally, it should become integrated in standard proteomic workflows as an add-on to conventional database search engines, which then would be able to provide improved identification.

Authors

  1. Rui Vitorino · University of Aveiro, University of Porto
  2. Sofia Guedes · University of Aveiro
  3. Fabio Trindade · University of Porto
  4. Inês Correia · University of Aveiro
  5. Gabriela Moura · University of Aveiro
  6. Paulo C. Carvalho · Carlos Chagas Institute, Fiocruz Paraná, University of California San Diego
  7. Manuel A. S. Santos · University of Aveiro
  8. Francisco Amado · University of Aveiro

Methods and tools

  • De novo protein sequencing review (Vitorino): Expert Review of Proteomics survey of de novo sequencing at the protein level: how MS/MS-derived peptide sequences are assembled into full protein sequences, the algorithmic and instrumental limits involved, and where the approach beats database search for organisms and proteoforms absent from reference databases.

Cites (28)

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