A Strong Cation Exchange Chromatography Protocol for Examining N-Terminal Proteoforms.

N-terminal acetylation N-terminal acetyltransferases N-terminomics

Journal

Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969

Informations de publication

Date de publication:
2022
Historique:
entrez: 6 5 2022
pubmed: 7 5 2022
medline: 11 5 2022
Statut: ppublish

Résumé

Especially in eukaryotes, the N-terminal acetylation status of a protein reveals translation initiation sites and substrate specificities and activities of N-terminal acetyltransferases (NATs). Here, we discuss a bottom-up proteomics protocol for the enrichment of N-terminal peptides via strong cation exchange chromatography. This protocol is based on depleting internal tryptic peptides from proteome digests through their retention on strong cation exchangers, leaving N-terminally acetylated/blocked peptides enriched among the nonretained peptides. As such, one can identify novel N-terminal proteoforms and quantify the degree of N-terminal protein acetylation.

Identifiants

pubmed: 35524124
doi: 10.1007/978-1-0716-2257-5_17
doi:

Substances chimiques

Peptides 0
Proteome 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

293-309

Informations de copyright

© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

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Auteurs

Esperanza Fernández (E)

VIB Center for Medical Biotechnology, Ghent, Belgium.
Department of Biomolecular Medicine, Ghent University, Ghent, Belgium.

Annelies Bogaert (A)

VIB Center for Medical Biotechnology, Ghent, Belgium.
Department of Biomolecular Medicine, Ghent University, Ghent, Belgium.

Evy Timmerman (E)

VIB Center for Medical Biotechnology, Ghent, Belgium.
Department of Biomolecular Medicine, Ghent University, Ghent, Belgium.
VIB Proteomics Core, Ghent, Belgium.

An Staes (A)

VIB Center for Medical Biotechnology, Ghent, Belgium.
Department of Biomolecular Medicine, Ghent University, Ghent, Belgium.
VIB Proteomics Core, Ghent, Belgium.

Francis Impens (F)

VIB Center for Medical Biotechnology, Ghent, Belgium.
Department of Biomolecular Medicine, Ghent University, Ghent, Belgium.
VIB Proteomics Core, Ghent, Belgium.

Kris Gevaert (K)

VIB Center for Medical Biotechnology, Ghent, Belgium. kris.gevaert@vib-ugent.be.
Department of Biomolecular Medicine, Ghent University, Ghent, Belgium. kris.gevaert@vib-ugent.be.

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