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...

Full description

Saved in:
Bibliographic Details
Published in:Methods in molecular biology (Clifton, N.J.) Vol. 2228; p. 225
Main Authors: Holtz, Anja, Basisty, Nathan, Schilling, Birgit
Format: Journal Article
Language:English
Published: United States 2021
Subjects:
Acetylation
Animals
Chromatography, High Pressure Liquid
Chromatography, Reverse-Phase
High-Throughput Screening Assays
Liver - chemistry
Mice
Protein Processing, Post-Translational
Proteins - analysis
Proteomics
Research Design
Spectrometry, Mass, Electrospray Ionization
Tandem Mass Spectrometry
Post-translational modifications
Acetylation
Quantification
Mass spectrometry
Data-independent acquisition
ISSN:1940-6029, 1940-6029
Online Access:Get more information
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:1940-6029
1940-6029
DOI:10.1007/978-1-0716-1024-4_16