Quantification and Identification of Post-Translational Modifications Using Modern Proteomics Approaches

Post-translational modifications (PTMs) occur dynamically, allowing cells to quickly respond to changes in the environment. Lysine residues can be targeted by several modifications including acylations (acetylation, succinylation, malonylation, glutarylation, and others), methylation, ubiquitination...

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Autores principales: Holtz, Anja, Basisty, Nathan, Schilling, Birgit
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2021
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8710235/
https://www.ncbi.nlm.nih.gov/pubmed/33950494
http://dx.doi.org/10.1007/978-1-0716-1024-4_16
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author Holtz, Anja
Basisty, Nathan
Schilling, Birgit
author_facet Holtz, Anja
Basisty, Nathan
Schilling, Birgit
author_sort Holtz, Anja
collection PubMed
description Post-translational modifications (PTMs) occur dynamically, allowing cells to quickly respond to changes in the environment. Lysine residues can be targeted by several modifications including acylations (acetylation, succinylation, malonylation, glutarylation, and others), methylation, ubiquitination, and other modifications. One of the most efficient methods for the identification of post-translational modifications is utilizing immunoaffinity enrichment followed by high-resolution mass spectrometry. This workflow can be coupled with comprehensive data-independent acquisition (DIA) mass spectrometry to be a high-throughput, label-free PTM quantification approach. Below we describe a detailed protocol to process tissue by homogenization and proteolytically digest proteins, followed by immunoaffinity enrichment of lysine-acetylated peptides to identify and quantify relative changes of acetylation comparing different conditions.
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spelling pubmed-87102352021-12-26 Quantification and Identification of Post-Translational Modifications Using Modern Proteomics Approaches Holtz, Anja Basisty, Nathan Schilling, Birgit Methods Mol Biol Article Post-translational modifications (PTMs) occur dynamically, allowing cells to quickly respond to changes in the environment. Lysine residues can be targeted by several modifications including acylations (acetylation, succinylation, malonylation, glutarylation, and others), methylation, ubiquitination, and other modifications. One of the most efficient methods for the identification of post-translational modifications is utilizing immunoaffinity enrichment followed by high-resolution mass spectrometry. This workflow can be coupled with comprehensive data-independent acquisition (DIA) mass spectrometry to be a high-throughput, label-free PTM quantification approach. Below we describe a detailed protocol to process tissue by homogenization and proteolytically digest proteins, followed by immunoaffinity enrichment of lysine-acetylated peptides to identify and quantify relative changes of acetylation comparing different conditions. 2021 /pmc/articles/PMC8710235/ /pubmed/33950494 http://dx.doi.org/10.1007/978-1-0716-1024-4_16 Text en https://creativecommons.org/licenses/by/4.0/Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ), which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the chapter’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.
spellingShingle Article
Holtz, Anja
Basisty, Nathan
Schilling, Birgit
Quantification and Identification of Post-Translational Modifications Using Modern Proteomics Approaches
title Quantification and Identification of Post-Translational Modifications Using Modern Proteomics Approaches
title_full Quantification and Identification of Post-Translational Modifications Using Modern Proteomics Approaches
title_fullStr Quantification and Identification of Post-Translational Modifications Using Modern Proteomics Approaches
title_full_unstemmed Quantification and Identification of Post-Translational Modifications Using Modern Proteomics Approaches
title_short Quantification and Identification of Post-Translational Modifications Using Modern Proteomics Approaches
title_sort quantification and identification of post-translational modifications using modern proteomics approaches
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8710235/
https://www.ncbi.nlm.nih.gov/pubmed/33950494
http://dx.doi.org/10.1007/978-1-0716-1024-4_16
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