Lysine Acetylation Stoichiometry Analysis at the Proteome Level
Gil, Jeovanis LU
and Encarnación-Guevara, Sergio
(2022)
In Methods in Molecular Biology
2420.
p.73-86
- Abstract
Lysine acetylation is a widespread posttranslational modification (PTM) in all kingdoms of live. A large number of proteins involved in most of biological pathways are targets of this PTM. The lysine acetylation is a reversible modification controlled by two main groups of enzymes, lysine acetyltransferases responsible for transferring the acetyl group of acetylCoA to the side chain of lysine residues and lysine deacetylases which effectively remove the acetyl tag. Dysregulation of enzymes that control acetylation and/or target proteins have been associated with a growing number of human pathologies. Lysine acetylation is largely a modification that occurs at low stoichiometry at its target sites. Here we describe a method to identify... (More)
Lysine acetylation is a widespread posttranslational modification (PTM) in all kingdoms of live. A large number of proteins involved in most of biological pathways are targets of this PTM. The lysine acetylation is a reversible modification controlled by two main groups of enzymes, lysine acetyltransferases responsible for transferring the acetyl group of acetylCoA to the side chain of lysine residues and lysine deacetylases which effectively remove the acetyl tag. Dysregulation of enzymes that control acetylation and/or target proteins have been associated with a growing number of human pathologies. Lysine acetylation is largely a modification that occurs at low stoichiometry at its target sites. Here we describe a method to identify lysine acetylation sites and estimate their site occupancy at the proteome scale. The method relies on a high-resolution mass spectrometry-based proteomics approach, which includes a specific chemical acetylation reaction on unmodified lysine residues that carry heavy isotopes. The procedures described here have been applied to cell line cultures and to clinically relevant samples stored as both snap-frozen and formalin-fixed paraffin-embedded (FFPE) tissues.
(Less)
- author
- Gil, Jeovanis
LU
and Encarnación-Guevara, Sergio
- organization
- publishing date
- 2022
- type
- Chapter in Book/Report/Conference proceeding
- publication status
- published
- subject
- keywords
- Cell lines, FFPE tissue, Frozen tissue, Lysine acetylation, Mass spectrometry-based proteomics, N-acetoxysuccinimide-d3, Stoichiometry
- host publication
- Methods in Molecular Biology
- series title
- Methods in Molecular Biology
- volume
- 2420
- pages
- 14 pages
- publisher
- Humana Press
- external identifiers
-
- pmid:34905167
- scopus:85121709547
- ISSN
- 1064-3745
- 1940-6029
- DOI
- 10.1007/978-1-0716-1936-0_7
- language
- English
- LU publication?
- yes
- id
- 25eaed71-39ec-4d69-be63-403d1212d6e3
- date added to LUP
- 2022-03-04 08:55:44
- date last changed
- 2025-03-10 11:32:10
@inbook{25eaed71-39ec-4d69-be63-403d1212d6e3,
abstract = {{<p>Lysine acetylation is a widespread posttranslational modification (PTM) in all kingdoms of live. A large number of proteins involved in most of biological pathways are targets of this PTM. The lysine acetylation is a reversible modification controlled by two main groups of enzymes, lysine acetyltransferases responsible for transferring the acetyl group of acetylCoA to the side chain of lysine residues and lysine deacetylases which effectively remove the acetyl tag. Dysregulation of enzymes that control acetylation and/or target proteins have been associated with a growing number of human pathologies. Lysine acetylation is largely a modification that occurs at low stoichiometry at its target sites. Here we describe a method to identify lysine acetylation sites and estimate their site occupancy at the proteome scale. The method relies on a high-resolution mass spectrometry-based proteomics approach, which includes a specific chemical acetylation reaction on unmodified lysine residues that carry heavy isotopes. The procedures described here have been applied to cell line cultures and to clinically relevant samples stored as both snap-frozen and formalin-fixed paraffin-embedded (FFPE) tissues.</p>}},
author = {{Gil, Jeovanis and Encarnación-Guevara, Sergio}},
booktitle = {{Methods in Molecular Biology}},
issn = {{1064-3745}},
keywords = {{Cell lines; FFPE tissue; Frozen tissue; Lysine acetylation; Mass spectrometry-based proteomics; N-acetoxysuccinimide-d3; Stoichiometry}},
language = {{eng}},
pages = {{73--86}},
publisher = {{Humana Press}},
series = {{Methods in Molecular Biology}},
title = {{Lysine Acetylation Stoichiometry Analysis at the Proteome Level}},
url = {{http://dx.doi.org/10.1007/978-1-0716-1936-0_7}},
doi = {{10.1007/978-1-0716-1936-0_7}},
volume = {{2420}},
year = {{2022}},
}