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Makarov, Dmytro; Telek, Andras; Becker, Tobias; Wrisberg, Marie-Kristin; Schneider, Sabine und Kielkowski, Pavel (2022): Clickable report tags for identification of modified peptides by mass spectrometry. In: Journal of Mass Spectrometry, Bd. 57, Nr. 3 [PDF, 2MB]

Abstract

The identification and quantification of modified peptides are critical for the functional characterization of post-translational protein modifications (PTMs) to elucidate their biological function. Nowadays, quantitative mass spectrometry coupled with various bioinformatic pipelines has been successfully used for the determination of a wide range of PTMs. However, direct characterization of low abundant protein PTMs in bottom-up proteomic workflow remains challenging. Here, we present the synthesis and evaluation of tandem mass spectrometry tags (TMT) which are introduced via click-chemistry into peptides bearing alkyne handles. The fragmentation properties of the two mass tags were validated and used for screening in a model system and analysis of AMPylated proteins. The presented tags provide a valuable tool for diagnostic peak generation to increase confidence in the identification of modified peptides and potentially for direct peptide-PTM quantification from various experimental conditions.

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