Exopeptidase Assisted N- and C-Terminal Proteome Sequencing
| Autor: | Andreas O. Helbig, Andreas Tholey |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Proteomics
Proteome biology Chemistry 010401 analytical chemistry Translation (biology) Saccharomyces cerevisiae Computational biology Exopeptidase 010402 general chemistry 01 natural sciences Genome Mass Spectrometry 0104 chemical sciences Analytical Chemistry Terminal (electronics) Exopeptidases biology.protein Protein Processing Post-Translational |
| Zdroj: | Analytical Chemistry. 92:5023-5032 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/acs.analchem.9b05288 |
| Popis: | Due to mechanisms such as proteolytic processing or alternative translation starts, in vivo proteoforms do not necessarily correspond directly to those encoded in the genome. Therefore, the knowledge of protein termini is an indispensable prerequisite to understand protein functions. So far, sequencing of protein N- and C-termini has been limited to single purified protein species, while the proteome-wide identification of N- and C-termini relies on the generation of single, terminal proteotypic peptides followed by chemical enrichment or depletion strategies to facilitate their detection via mass spectrometry (MS). To overcome the numerous limitations in such approaches, we present an alternative concept that readily enables unbiased ladder sequencing of protein N- and C-termini. The approach combines exopeptidase digestions of the proteome with two-dimensional chromatographic separation and tandem-MS. We demonstrate the potential of the methodology by analyzing the N- and C-terminome of |
| Databáze: | OpenAIRE |
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