A Combined Chemical Derivatization/Mass Spectrometric Method for the Enhanced Detection and Relative Quantification of Protein Ubiquitination

Mass spectrometry (MS) is a sensitive analytical technique with wide application across the sciences including for the detection of peptides and proteins in biological analysis. Ubiquitinated (Ub) proteins are typically analyzed by proteolytic digestion and subsequent chromatographic separation foll...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 1977; p. 17
Main Authors Chicooree, Navin, Griffiths, John R
Format Journal Article
LanguageEnglish
Published United States 2019
Subjects
Data Interpretation, Statistical
Gas Chromatography-Mass Spectrometry
Mass Spectrometry - methods
Protein Processing, Post-Translational
Proteomics - methods
Staining and Labeling
Ubiquitinated Proteins - chemistry
Workflow
Diagnostic ions
Ubiquitin
SWATH
Posttranslational modifications
Mass spectrometry
Data-independent acquisition
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Summary:Mass spectrometry (MS) is a sensitive analytical technique with wide application across the sciences including for the detection of peptides and proteins in biological analysis. Ubiquitinated (Ub) proteins are typically analyzed by proteolytic digestion and subsequent chromatographic separation followed by MS detection of the resulting isopeptides. Here we describe a novel method which enables enhanced detection of this important posttranslational modification (PTM) by use of a simple chemical labeling strategy prior to Data-Independent Acquisition (DIA) using a SWATH-based acquisition approach on a suitable Quadrupole-Time-Of-Flight (Q-TOF) mass spectrometer.
ISSN:1940-6029
DOI:10.1007/978-1-4939-9232-4_2