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Identifying transglutaminase reaction products via mass spectrometry as exemplified by the MUC2 mucin - Pitfalls and traps

Journal article
Authors Liisa Arike
Gunnar C. Hansson
Christian V Recktenwald
Published in Analytical Biochemistry
Volume 597
ISSN 0003-2697
Publication year 2020
Published at Institute of Biomedicine, Department of Medical Biochemistry and Cell Biology
Language en
Links dx.doi.org/10.1016/j.ab.2020.113668
Keywords Mucus, MUC2, Isopeptide bond, Cross-link, Transglutaminase, identification, peptides, deamidation, glutamine, ms/ms, Biochemistry & Molecular Biology, Chemistry
Subject categories Biochemistry and Molecular Biology

Abstract

In order to demonstrate transglutaminase activity in biological samples immunological as well as glutamine- and amine-donor based assays are commonly used. However, the identification of the transglutaminase reaction product, i. e. the isopeptide cross-linked peptides/proteins or the deamidated protein/peptide are often neglected. This article describes a workflow for the detection of the products of transglutaminase-catalyzed reactions. In particular, possible pitfalls and traps that can arise during the mass spectrometry-based identification of isopeptide cross-links are addressed and characterised on actual samples.

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